Product Citations: 238

Fig.1.G showing Western Blotting from the publication: Disruption of RCAN1.4 expression mediated by YY1/HDAC2 modulates chronic renal allograft interstitial fibrosis.
Fig.2.G showing Western Blotting from the publication: Disruption of RCAN1.4 expression mediated by YY1/HDAC2 modulates chronic renal allograft interstitial fibrosis.
Fig.2.A showing Western Blotting from the publication: Disruption of RCAN1.4 expression mediated by YY1/HDAC2 modulates chronic renal allograft interstitial fibrosis.
Fig.2.D showing Western Blotting from the publication: Disruption of RCAN1.4 expression mediated by YY1/HDAC2 modulates chronic renal allograft interstitial fibrosis.
Fig.3.D showing Western Blotting from the publication: Disruption of RCAN1.4 expression mediated by YY1/HDAC2 modulates chronic renal allograft interstitial fibrosis.
Fig.5.A showing Western Blotting from the publication: Disruption of RCAN1.4 expression mediated by YY1/HDAC2 modulates chronic renal allograft interstitial fibrosis.
Fig.5.E showing Western Blotting from the publication: Disruption of RCAN1.4 expression mediated by YY1/HDAC2 modulates chronic renal allograft interstitial fibrosis.
Fig.5.B showing Western Blotting from the publication: A CRISPR/Cas9-Based Assay for High-Throughput Studies of Cancer-Induced Innervation.
Fig.3.E showing Western Blotting from the publication: RSPO2 promotes progression of ovarian cancer through dual receptor-mediated FAK/Src signaling activation.
Fig.4.F showing Western Blotting from the publication: RSPO2 promotes progression of ovarian cancer through dual receptor-mediated FAK/Src signaling activation.
Fig.5.A showing Western Blotting from the publication: CARMA3 Promotes Colorectal Cancer Cell Motility and Cancer Stemness via YAP-Mediated NF-κB Activation.
Fig.2.H showing Western Blotting from the publication: Protective function of interleukin-22 in pulmonary fibrosis.
Fig.4.N showing Western Blotting from the publication: Protective function of interleukin-22 in pulmonary fibrosis.
Fig.7.A showing Western Blotting from the publication: Arginine methylation of SHANK2 by PRMT7 promotes human breast cancer metastasis through activating endosomal FAK signalling.
Fig.5.A showing Western Blotting from the publication: Fibroblast-enriched endoplasmic reticulum protein TXNDC5 promotes pulmonary fibrosis by augmenting TGFβ signaling through TGFBR1 stabilization.
Fig.7.D showing Western Blotting from the publication: Arginine methylation of SHANK2 by PRMT7 promotes human breast cancer metastasis through activating endosomal FAK signalling.
Fig.7.G showing Western Blotting from the publication: Arginine methylation of SHANK2 by PRMT7 promotes human breast cancer metastasis through activating endosomal FAK signalling.
Fig.7.J showing Western Blotting from the publication: Arginine methylation of SHANK2 by PRMT7 promotes human breast cancer metastasis through activating endosomal FAK signalling.
Fig.3.A showing Western Blotting from the publication: Arginine methylation of SHANK2 by PRMT7 promotes human breast cancer metastasis through activating endosomal FAK signalling.
Fig.4.E showing Western Blotting from the publication: Low levels of AMPK promote epithelial-mesenchymal transition in lung cancer primarily through HDAC4- and HDAC5-mediated metabolic reprogramming.
Fig.4.F showing Western Blotting from the publication: Low levels of AMPK promote epithelial-mesenchymal transition in lung cancer primarily through HDAC4- and HDAC5-mediated metabolic reprogramming.
Fig.5.H showing Western Blotting from the publication: Low levels of AMPK promote epithelial-mesenchymal transition in lung cancer primarily through HDAC4- and HDAC5-mediated metabolic reprogramming.
Fig.5.D showing Western Blotting in a Homo sapiens (Human) sample from the publication: Inhibition of TGF-β signalling in combination with nal-IRI plus 5-Fluorouracil/Leucovorin suppresses invasion and prolongs survival in pancreatic tumour mouse models.
Fig.2.C showing Western Blotting in a Homo sapiens (Human) sample from the publication: Inhibition of TGF-β signalling in combination with nal-IRI plus 5-Fluorouracil/Leucovorin suppresses invasion and prolongs survival in pancreatic tumour mouse models.
Fig.3.A showing Western Blotting from the publication: Quercetin Inhibits Cell Survival and Metastatic Ability via the EMT-mediated Pathway in Oral Squamous Cell Carcinoma.
41 images found

Methylglyoxal-Stimulated Mesothelial Cells Prompted Fibroblast-to-Proto-Myofibroblast Transition.

In International Journal of Molecular Sciences on 19 January 2025 by Wei, Y. S., Tsai, S. Y., et al.

During long-term peritoneal dialysis, peritoneal fibrosis (PF) often happens and results in ultrafiltration failure, which directly leads to the termination of dialysis. The accumulation of extracellular matrix produced from an increasing number of myofibroblasts was a hallmark characteristic of PF. To date, glucose degradation products (GDPs, i.e., methylglyoxal (MGO)) that appeared during the heating and storage of the dialysate are considered to be key components to initiating PF, but how GDPs lead to the activation of myofibroblast in fibrotic peritoneum has not yet been fully elucidated. In this study, mesothelial cell line (MeT-5A) and fibroblast cell line (MRC-5) were used to investigate the transcriptomic and proteomic changes to unveil the underlying mechanism of MGO-induced PF. Our transcriptomic data from the MGO-stimulated mesothelial cells showed upregulation of genes involved in pro-inflammatory, apoptotic, and fibrotic pathways. While no phenotypic changes were noted on fibroblasts after direct MGO, supernatant from MGO-stimulated mesothelial cells promoted fibroblasts to change into proto-myofibroblasts, activated fibroblasts in the first stage toward myofibroblasts. In conclusion, this study showed that MGO-stimulated mesothelial cells promoted fibroblast-to-proto-myofibroblast transition; however, additional involvement of other factors or cells (e.g., macrophages) may be needed to complete the transformation into myofibroblasts.

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Extracellular Vesicles from Human Induced Pluripotent Stem Cells Exhibit a Unique MicroRNA and CircRNA Signature.

In International Journal of Biological Sciences on 12 December 2024 by Barilani, M., Peli, V., et al.

Extracellular vesicles (EV) have emerged as promising cell-free therapeutics in regenerative medicine. However, translating primary cell line-derived EV to clinical applications requires large-scale manufacturing and several challenges, such as replicative senescence, donor heterogeneity, and genetic instability. To address these limitations, we used a reprogramming approach to generate human induced pluripotent stem cells (hiPSC) from the young source of cord blood mesenchymal stem/stromal cells (CBMSC). Capitalizing on their inexhaustible supply potential, hiPSC offer an attractive EV reservoir. Our approach encompassed an exhaustive characterization of hiPSC-EV, aligning with the rigorous MISEV2023 guidelines. Analyses demonstrated physical features compatible with small EV (sEV) and established their identity and purity. Moreover, the sEV-shuttled non-coding (nc) RNA landscape, focusing on the microRNA and circular RNA cargo, completed the molecular signature. The kinetics of the hiPSC-sEV release and cell internalization assays unveiled robust EV production and consistent uptake by human neurons. Furthermore, hiPSC-sEV demonstrated ex vivo cell tissue-protective properties. Finally, via bioinformatics, the potential involvement of the ncRNA cargo in the hiPSC-sEV biological effects was explored. This study significantly advances the understanding of pluripotent stem cell-derived EV. We propose cord blood MSC-derived hiPSC as a promising source for potentially therapeutic sEV.
© The author(s).

  • Stem Cells and Developmental Biology
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PKMζ, a Brain-specific PKCζ Isoform, is Required for Glycolysis and Myofibroblastic Activation of Hepatic Stellate Cells.

In Cellular and Molecular Gastroenterology and Hepatology on 15 November 2024 by Wang, X., Wang, Y., et al.

Transforming growth factor (TGF)β1 induces plasma membrane (PM) accumulation of glucose transporter 1 (Glut1) required for glycolysis of hepatic stellate cells (HSCs) and HSC activation. This study aimed to understand how Glut1 is anchored/docked onto the PM of HSCs.HSC expression of protein kinase M zeta isoform (PKMζ) was detected by reverse transcription polymerase chain reaction (RT-PCR), Western blotting, and immunofluorescence. PKMζ level was manipulated by short hairpin RNA (shRNA) or overexpression; HSC activation was assessed by cell expression of activation markers; PM Glut1, glucose uptake, and glycolysis of HSCs were analyzed by biotinylation, 2-NBDG-based assay, and Seahorse Glycolysis Stress Test. Phospho-mutants of vasodilator-stimulated phosphorylated protein (VASP) were created by site-directed mutagenesis. TGFβ transcriptome was obtained by RNA sequencing. Single-cell RNA sequencing datasets and immunofluorescence were leveraged to analyze PKMζ expression in cancer-associated fibroblasts (CAFs) of colorectal liver metastases. Function of HSC PKMζ was determined by tumor/HSC co-implantation study.Primary human and murine HSCs express PKMζ, but not full-length PKCζ. PKMζ knockdown suppresses, whereas PKMζ overexpression potentiates PM accumulation of Glut1, glycolysis, and HSC activation induced by TGFβ1. Mechanistically, PKMζ binds to and induces VASP phosphorylation at serines 157 and 239 facilitating anchoring/docking of Glut1 onto the PM of HSCs. PKMζ expression is increased in the CAFs of murine and patient colorectal liver metastases compared with quiescent HSCs. Targeting PKMζ suppresses transcriptome, CAF activation of HSCs, and colorectal tumor growth in mice.Because HSCs are also a major contributor of liver fibrosis, our data highlight PKMζ and VASP as targets to inhibit metabolic reprogramming, HSC activation, liver fibrosis, and the pro-metastatic microenvironment of the liver.Copyright © 2025 The Authors. Published by Elsevier Inc. All rights reserved.

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Primary cilia promote EMT-induced triple-negative breast tumor heterogeneity and resistance to therapy

Preprint on BioRxiv : the Preprint Server for Biology on 14 September 2024 by Tessier, C., Derrien, J., et al.

Tumor heterogeneity and plasticity, driven by Epithelial-Mesenchymal Transition (EMT), enable cancer therapeutic resistance. We previously showed that EMT promotes primary cilia formation, which enables stemness and tumorigenesis in triple-negative breast cancer (TNBC). Here, we establish a role for primary cilia in human TNBC chemotherapeutic resistance. We developed patient-derived organoids, and showed that these recapitulated the cellular heterogeneity of TNBC biopsies. Notably, one of the identified cell states bore a quasi-mesenchymal phenotype, primary cilia, and stemness signatures. We treated our TNBC organoids with chemotherapeutics and observed partial killing. The surviving cells with organoid-reconstituting capacity showed selective enrichment for the quasi-mesenchymal ciliated cell subpopulation. Genomic analyses argue that this enrichment reflects a combination of pre-existing cells and ones that arose through drug-induced cellular plasticity. We developed a family of small-molecule inhibitors of ciliogenesis and show that these, or genetic ablation of primary cilia, suppress chemoresistance. We conclude that primary cilia help TNBC to evade chemotherapy.

  • Homo sapiens (Human)
  • Cancer Research
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Novel high throughput 3D ECM remodeling assay identifies MEK as key driver of fibrotic fibroblast activity

Preprint on BioRxiv : the Preprint Server for Biology on 27 August 2024 by Liao, C. Y., Hundscheid, J. H., et al.

ABSTRACT In fibrotic tissues, activated fibroblasts remodel the collagen-rich extracellular matrix (ECM). Intervening with this process represents a candidate therapeutic strategy to attenuate disease progression. Models that generate quantitative data on 3D fibroblast-mediated ECM remodeling with the reproducibility and throughput needed for drug testing are lacking. Here, we develop a model that fits this purpose and produces combined quantitative information on drug efficacy and cytotoxicity. We use microinjection robotics to design patterns of fibrillar collagen-embedded fibroblast clusters and apply automated microscopy and image analysis to quantify ECM remodeling between-, and cell viability within clusters of TGFβ-activated primary human skin or lung fibroblasts. We apply this assay to compound screening and reveal actionable targets to suppress fibrotic ECM remodeling. Strikingly, we find that after an initial phase of fibroblast activation by TGFβ, canonical TGFβ signaling is dispensable and, instead, non-canonical activation of MEK-ERK signaling drives ECM remodeling. Moreover, we reveal that higher concentrations of two TGFβ receptor inhibitors while blocking canonical TGFβ signaling, in fact stimulate this MEK-mediated profibrotic ECM remodeling activity.

  • Homo sapiens (Human)
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Disruption of RCAN1.4 expression mediated by YY1/HDAC2 modulates chronic renal allograft interstitial fibrosis.

In Cell Death Discov on 28 July 2023 by Zhang, J., Zhang, Y., et al.

Fig.1.G
Fig.1.G showing Western Blotting from the publication: Disruption of RCAN1.4 expression mediated by YY1/HDAC2 modulates chronic renal allograft interstitial fibrosis.
  • WB
RCAN1.4 expression was down-regulated in vivo and in vitro.The mouse renal transplanted models of chronic renal allograft interstitial fibrosis were established as described in methods and materials. A Schematic diagram of mouse experimental process. B HE, Masson trichrome staining and Sirius Red... more
RCAN1.4 expression was down-regulated in vivo and in vitro.The mouse renal transplanted models of chronic renal allograft interstitial fibrosis were established as described in methods and materials. A Schematic diagram of mouse experimental process. B HE, Masson trichrome staining and Sirius Red staining stains of kidney interstitium tissue of the two groups of mice (×100, scale bar, 100 μm). Renal parenchyma exhibiting tubular dilation (black *) and ECM deposition in interstitial space of kidney (black # and &). C Immunohistochemistry signals of RCAN1, YY1, E-cad, α-SMA, and TNF-α. D Tubulointerstitial damage scores and the proportion of Masson’s positive area were assessed in different groups (n = 6, *P < 0.05). E Statistical analysis of the protein’s positive area was performed in different groups (n = 6, *P < 0.05). F Western blotting analysis was performed after collecting kidney lysates. Representative blots of RCAN1.4 and YY1(n = 6). In vivo, TNF-α(0–100 ng/mL) was applied to HK‐2 cells to induce EMT. (G-H) Representative blots of RCAN1.4, YY1, EMT markers (COL1, FN, E-cad and α-SMA) (n = 3). I Co-localization of RCAN1 and YY1 protein expression in TNF-α-stimulated HK-2 cells was examined by immunofluorescence (scale bars, 100 μm). J The mRNA levels of TNF-α were detected by qPCR (n = 3). The results are expressed as the mean ± standard error of the mean (SEM). for 3–4 independent experiments. *p < 0.05, as indicated. less

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Disruption of RCAN1.4 expression mediated by YY1/HDAC2 modulates chronic renal allograft interstitial fibrosis.

In Cell Death Discov on 28 July 2023 by Zhang, J., Zhang, Y., et al.

Fig.2.G
Fig.2.G showing Western Blotting from the publication: Disruption of RCAN1.4 expression mediated by YY1/HDAC2 modulates chronic renal allograft interstitial fibrosis.
  • WB
RCAN1.4-mediated suppressive function on EMT induced by TNF-α through modulating reactive species in vitro.HK-2 cells were transfected with RV230-RCAN1.4 plasmid or RCAN1.4-RNAi to change the expression of RCAN1.4, and treated with H2O2 or NAC with indicated concentration, followed by TNF-α stimu... more
RCAN1.4-mediated suppressive function on EMT induced by TNF-α through modulating reactive species in vitro.HK-2 cells were transfected with RV230-RCAN1.4 plasmid or RCAN1.4-RNAi to change the expression of RCAN1.4, and treated with H2O2 or NAC with indicated concentration, followed by TNF-α stimuli. A, D, G, I Representative blots of EMT markers (COL1, FN, E-cad, and α-SMA) (n = 3). B, E, H, J Representative blots of anti-oxidant proteins (SOD1, SOD2, and Catalase) (n = 3). C, F O2⋅− levels were detected by HE probe and were quantified(n = 3). The results are expressed as the mean ± standard error of the mean (SEM). for 3–4 independent experiments. *p < 0.05, **p < 0.05, as indicated. less

Collected and cropped from Cell Death Discov by CiteAb, provided under a CC-BY license

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Disruption of RCAN1.4 expression mediated by YY1/HDAC2 modulates chronic renal allograft interstitial fibrosis.

In Cell Death Discov on 28 July 2023 by Zhang, J., Zhang, Y., et al.

Fig.2.A
Fig.2.A showing Western Blotting from the publication: Disruption of RCAN1.4 expression mediated by YY1/HDAC2 modulates chronic renal allograft interstitial fibrosis.
  • WB
RCAN1.4-mediated suppressive function on EMT induced by TNF-α through modulating reactive species in vitro.HK-2 cells were transfected with RV230-RCAN1.4 plasmid or RCAN1.4-RNAi to change the expression of RCAN1.4, and treated with H2O2 or NAC with indicated concentration, followed by TNF-α stimu... more
RCAN1.4-mediated suppressive function on EMT induced by TNF-α through modulating reactive species in vitro.HK-2 cells were transfected with RV230-RCAN1.4 plasmid or RCAN1.4-RNAi to change the expression of RCAN1.4, and treated with H2O2 or NAC with indicated concentration, followed by TNF-α stimuli. A, D, G, I Representative blots of EMT markers (COL1, FN, E-cad, and α-SMA) (n = 3). B, E, H, J Representative blots of anti-oxidant proteins (SOD1, SOD2, and Catalase) (n = 3). C, F O2⋅− levels were detected by HE probe and were quantified(n = 3). The results are expressed as the mean ± standard error of the mean (SEM). for 3–4 independent experiments. *p < 0.05, **p < 0.05, as indicated. less

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Disruption of RCAN1.4 expression mediated by YY1/HDAC2 modulates chronic renal allograft interstitial fibrosis.

In Cell Death Discov on 28 July 2023 by Zhang, J., Zhang, Y., et al.

Fig.2.D
Fig.2.D showing Western Blotting from the publication: Disruption of RCAN1.4 expression mediated by YY1/HDAC2 modulates chronic renal allograft interstitial fibrosis.
  • WB
RCAN1.4-mediated suppressive function on EMT induced by TNF-α through modulating reactive species in vitro.HK-2 cells were transfected with RV230-RCAN1.4 plasmid or RCAN1.4-RNAi to change the expression of RCAN1.4, and treated with H2O2 or NAC with indicated concentration, followed by TNF-α stimu... more
RCAN1.4-mediated suppressive function on EMT induced by TNF-α through modulating reactive species in vitro.HK-2 cells were transfected with RV230-RCAN1.4 plasmid or RCAN1.4-RNAi to change the expression of RCAN1.4, and treated with H2O2 or NAC with indicated concentration, followed by TNF-α stimuli. A, D, G, I Representative blots of EMT markers (COL1, FN, E-cad, and α-SMA) (n = 3). B, E, H, J Representative blots of anti-oxidant proteins (SOD1, SOD2, and Catalase) (n = 3). C, F O2⋅− levels were detected by HE probe and were quantified(n = 3). The results are expressed as the mean ± standard error of the mean (SEM). for 3–4 independent experiments. *p < 0.05, **p < 0.05, as indicated. less

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Disruption of RCAN1.4 expression mediated by YY1/HDAC2 modulates chronic renal allograft interstitial fibrosis.

In Cell Death Discov on 28 July 2023 by Zhang, J., Zhang, Y., et al.

Fig.3.D
Fig.3.D showing Western Blotting from the publication: Disruption of RCAN1.4 expression mediated by YY1/HDAC2 modulates chronic renal allograft interstitial fibrosis.
  • WB
Recombinant adeno-associated virus-mediated overexpression of RCAN1.4 attenuated chronic renal allograft interstitial fibrosis.The mouse models were established as described in the “Materials and methods” section. A HE, Masson trichrome staining, and Sirius Red staining stains of kidney interstit... more
Recombinant adeno-associated virus-mediated overexpression of RCAN1.4 attenuated chronic renal allograft interstitial fibrosis.The mouse models were established as described in the “Materials and methods” section. A HE, Masson trichrome staining, and Sirius Red staining stains of kidney interstitium tissue of the two groups of mice (X400, Scale bar, 100 μm). Renal parenchyma exhibiting tubular dilation (black *) and ECM deposition in interstitial space of kidney (black # and &). B Immunohistochemistry signals of RCAN1, YY1, E-cad, and α-SMA. Statistical analysis of the protein’s positive area was performed in different groups (n = 6, *p < 0.05). C Renal O2⋅− levels were examined by DHE staining (scale bars, 100 μm). D Representative blots of RCAN1.4 and EMT markers (COL1, FN, E-cad, and α-SMA) (n = 6). E Representative blots of anti-oxidant proteins (SOD1, SOD2, and Catalase) (n = 6). Data represent the mean ± s.e.m. *p < 0.05, as indicated. less

Collected and cropped from Cell Death Discov by CiteAb, provided under a CC-BY license

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Disruption of RCAN1.4 expression mediated by YY1/HDAC2 modulates chronic renal allograft interstitial fibrosis.

In Cell Death Discov on 28 July 2023 by Zhang, J., Zhang, Y., et al.

Fig.5.A
Fig.5.A showing Western Blotting from the publication: Disruption of RCAN1.4 expression mediated by YY1/HDAC2 modulates chronic renal allograft interstitial fibrosis.
  • WB
YY1 modulated TNF-α-induced reactive species and EMT dependently on RCAN1.4.A, E Representative blots of EMT markers (COL1, FN, E-cad, and α-SMA) and GAPDH (n = 3). B, F Representative blots of RCAN1.4, YY1, and anti-oxidant proteins (SOD1, SOD2, and Catalase). C, D (G, H) O2⋅− levels were detect... more
YY1 modulated TNF-α-induced reactive species and EMT dependently on RCAN1.4.A, E Representative blots of EMT markers (COL1, FN, E-cad, and α-SMA) and GAPDH (n = 3). B, F Representative blots of RCAN1.4, YY1, and anti-oxidant proteins (SOD1, SOD2, and Catalase). C, D (G, H) O2⋅− levels were detected by HE probe, and were quantified (n = 3), *p < 0.05, as indicated. The results are expressed as the mean ± standard error of the mean (SEM) for 3–4 independent experiments. less

Collected and cropped from Cell Death Discov by CiteAb, provided under a CC-BY license

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Disruption of RCAN1.4 expression mediated by YY1/HDAC2 modulates chronic renal allograft interstitial fibrosis.

In Cell Death Discov on 28 July 2023 by Zhang, J., Zhang, Y., et al.

Fig.5.E
Fig.5.E showing Western Blotting from the publication: Disruption of RCAN1.4 expression mediated by YY1/HDAC2 modulates chronic renal allograft interstitial fibrosis.
  • WB
YY1 modulated TNF-α-induced reactive species and EMT dependently on RCAN1.4.A, E Representative blots of EMT markers (COL1, FN, E-cad, and α-SMA) and GAPDH (n = 3). B, F Representative blots of RCAN1.4, YY1, and anti-oxidant proteins (SOD1, SOD2, and Catalase). C, D (G, H) O2⋅− levels were detect... more
YY1 modulated TNF-α-induced reactive species and EMT dependently on RCAN1.4.A, E Representative blots of EMT markers (COL1, FN, E-cad, and α-SMA) and GAPDH (n = 3). B, F Representative blots of RCAN1.4, YY1, and anti-oxidant proteins (SOD1, SOD2, and Catalase). C, D (G, H) O2⋅− levels were detected by HE probe, and were quantified (n = 3), *p < 0.05, as indicated. The results are expressed as the mean ± standard error of the mean (SEM) for 3–4 independent experiments. less

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A CRISPR/Cas9-Based Assay for High-Throughput Studies of Cancer-Induced Innervation.

In Cancers (Basel) on 29 March 2023 by Galappaththi, S. L., Katz, B., et al.

Fig.5.B
Fig.5.B showing Western Blotting from the publication: A CRISPR/Cas9-Based Assay for High-Throughput Studies of Cancer-Induced Innervation.
  • WB
Breast cancer plasticity promotes neuron precursor axonogenesis. (A) To test the model developed for studying cancer-induced neuronal differentiation, we developed a model of breast cancer plasticity and epithelial-mesenchymal transition (EMT). The NMuMG cell cultures from ATCC are mostly epithel... more
Breast cancer plasticity promotes neuron precursor axonogenesis. (A) To test the model developed for studying cancer-induced neuronal differentiation, we developed a model of breast cancer plasticity and epithelial-mesenchymal transition (EMT). The NMuMG cell cultures from ATCC are mostly epithelial cells but are heterogenous cultures having mixed phenotypes, with a few cells exhibiting mesenchymal morphologies (red arrows). NMuMG clones with epithelial (NMuMG-E) and mesenchymal (NMuMG-M) morphologies were isolated by single-cell seeding and regrowing of isogenic culture having epithelial or mesenchymal characteristics. (B) Western blot analysis of the regenerated culture confirms the occurrence of an EMT, consisting of the expression of the epithelial marker E-cadherin in NMuMG-E cells that disappears in NMuMG-M cells and the acquisition of the expression of mesenchymal markers fibronectin, N-cadherin, and Slug in NMuMG-M cells (Original Western blot, see Supplementary File S1). GAPDH serves as a loading control. (C) Wound healing assay shows a greater velocity of migration in NMuMG-M cells compared to NMuMG-E cells, confirming the biological impact of the EMT on the cells’ aggressive behaviors. (D) To verify the relevance of NMuMG E/M to reflect the cell EMT-related plasticity functionally, we performed wound healing assays and quantified them by live-cell microscopy with the workflow published by Jonkman and colleagues [47]. (E) Fluorescence microscopy imaging of PC12-TUBB3-GFP cells in coculture with NMuMG-E or NMuMG-M constitutively expressing the mCherry fluorophore (red Channel), show increased GFP signal with the NMuMG-M cells, consistent with the development of extended neurites (White arrow). (F) PC12-TUBB-GFP cells were cultivated as unstimulated monoculture (Alone), incubated in direct coculture with either epithelial (+E cells) or mesenchymal (+M cells) cells, or treated with NGF (100 ng/mL) (+NGF) for 5 days as a positive control for differentiation. Flow cytometry revealed a moderate induction of PC12 neuronal precursor differentiation under NMuMG-E stimulation but a robust induction of differentiation under stimulation with NMuMG-M, where neuronal cells developed even more differentiation than those stimulated with NGF. (G) Cocultures of PC12-TUBB3-GFP-mCherry (constitutively expressing mCherry fluorophore) mixed with NMuMG-E or NMuMG-M cells were analyzed using an automated fluorescent plate reader. The mCherry expression allowed normalization of the assay by reflecting quantities of PC12 cells in the wells, and the mCherry/GFP ratio was used as an indicator of neuronal differentiation. Statistics: unpaired Student t-test ** p < 0.01; *** p < 0.001. Scale bar: 50 µm. less

Collected and cropped from Cancers (Basel) by CiteAb, provided under a CC-BY license

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RSPO2 promotes progression of ovarian cancer through dual receptor-mediated FAK/Src signaling activation.

In iScience on 21 October 2022 by Pan, R., Yu, Y., et al.

Fig.3.E
Fig.3.E showing Western Blotting from the publication: RSPO2 promotes progression of ovarian cancer through dual receptor-mediated FAK/Src signaling activation.
  • WB
Effects of RSPO2 on ovarian cancer cell adhesion, EMT, and cell cycle progression(A) GSEA of the TCGA ovarian cancer cohort showing that pathways associated with the cytoskeleton and focal adhesion were enriched in tumors with high RSPO2 expression. TCGA dataset was separated into two groups base... more
Effects of RSPO2 on ovarian cancer cell adhesion, EMT, and cell cycle progression(A) GSEA of the TCGA ovarian cancer cohort showing that pathways associated with the cytoskeleton and focal adhesion were enriched in tumors with high RSPO2 expression. TCGA dataset was separated into two groups based on the mRNA expression of RSPO2. The expression cutoff for KEGG analysis is first (RSPO2-High) and last quartile (RSPO2-Low) of mRNA expression of RSPO2.(B) Effect of RSPO2 on cell adhesion. Quantitation of adhesion and cell attachment, as evaluated by cellular binding to vitronectin, was performed using vector control cells (Vec) and cells stably overexpressing RSPO2 (RS2). ∗∗∗p < 0.001 vs. Vec.(C) Representative light micrographs showing that stable overexpression of RSPO2 altered the morphology of ovarian cancer cells (magnification, 200×).(D) Phalloidin staining showing actin filaments in A2780 and OVCAR3 cells stably overexpressing RSPO2. Scale bar, 20 μm.(E) Western blot analysis of EMT and cell cycle markers in cells stably overexpressing RSPO2.(F) Flow cytometric analysis of the cell cycle in RSPO2-silenced ovarian cancer cells. Cells were infected with lentivirus containing shRS2 or nontargeting shNC for 48 h.The data shown are the mean ± SD of a representative experiment performed in triplicate (lower panel). ∗∗p < 0.01 and p∗∗∗ <0.001 vs. shNC; two-tailed Student’s t-test. Error bars indicate mean ± SD. less

Collected and cropped from iScience by CiteAb, provided under a CC-BY license

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RSPO2 promotes progression of ovarian cancer through dual receptor-mediated FAK/Src signaling activation.

In iScience on 21 October 2022 by Pan, R., Yu, Y., et al.

Fig.4.F
Fig.4.F showing Western Blotting from the publication: RSPO2 promotes progression of ovarian cancer through dual receptor-mediated FAK/Src signaling activation.
  • WB
RSPO2 promotes ovarian cancer growth and metastasis through FAK/Src signaling cascades(A) KEGG enrichment analysis of the pathways affected by overexpression of RSPO2 in A2780 cells.(B) Effects of RSPO2 overexpression on the phosphorylation of FAK (Tyr397), Src (Tyr416, Y416), Src (Tyr527, Y527),... more
RSPO2 promotes ovarian cancer growth and metastasis through FAK/Src signaling cascades(A) KEGG enrichment analysis of the pathways affected by overexpression of RSPO2 in A2780 cells.(B) Effects of RSPO2 overexpression on the phosphorylation of FAK (Tyr397), Src (Tyr416, Y416), Src (Tyr527, Y527), EGFR (Tyr1068), and Akt (Ser473). Proteins were extracted from A2780 and OVCAR3 cells stably overexpressing RSPO2 (RS2, pooled) or empty vector (Vec) and were analyzed by Western blotting.(C) The effects of RSPO2 knockdown on the phosphorylation of FAK, Src, and Akt were analyzed by Western blotting. Cells were infected with lentivirus containing shRS2 or nontargeting shNC for 48 h.(D) Representative images (left panel) and quantification (right panel) of Transwell assay results showing that the FAK inhibitor defactinib (Def) abolished the promotive effect of RSPO2 overexpression on cell migration. Cells were treated with DMSO or 1.0 μM defactinib for 3 h.(E) Representative images (left panel) and quantification (right panel) of colony formation assay results showing that the Src inhibitor saracatinib (Sar) abolished the promotive effect of RSPO2 overexpression on cell proliferation. Cells were treated with DMSO or saracatinib (Sar, 10 μM) for 1 h.(F) Defactinib abolished the effect of RSPO2 on the expression of EMT markers and MMP7. A2780 and OVCAR3 cells stably overexpressing RSPO2 (RS2, pooled) or empty vector (Vec) were treated with DMSO or 1.0 μM defactinib for 3 h.(G) Saracatinib blocked RSPO2-induced EGFR/Akt phosphorylation.Cells were treated with DMSO or saracatinib (Sar, 10 μM) for 24 h n.s., not significant; p∗∗∗ <0.001; two-tailed Student’s t test. Error bars indicate mean ± SD. less

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CARMA3 Promotes Colorectal Cancer Cell Motility and Cancer Stemness via YAP-Mediated NF-κB Activation.

In Cancers (Basel) on 26 November 2021 by Chang, T. Y., Wu, C. T., et al.

Fig.5.A
Fig.5.A showing Western Blotting from the publication: CARMA3 Promotes Colorectal Cancer Cell Motility and Cancer Stemness via YAP-Mediated NF-κB Activation.
  • WB
CARMA3 induced EMT by upregulation of Slug. (A,B) The protein expression levels of indicated EMT-related proteins were determined and qualified in CARMA3-overexpressed and silenced cells by the Western blot assay (full WB in File S2). Results are shown as the mean ± S.D. of three independent expe... more
CARMA3 induced EMT by upregulation of Slug. (A,B) The protein expression levels of indicated EMT-related proteins were determined and qualified in CARMA3-overexpressed and silenced cells by the Western blot assay (full WB in File S2). Results are shown as the mean ± S.D. of three independent experiments. *, p < 0.05, compared to SW480/Ctrl or HCT116/shCtrl cells. (C,D) The protein expression levels of CARMA3 and Slug were analyzed by Western blot assay (full WB in File S2). α-Tubulin was used as the internal protein loading control. Each was performed in three independent experiments. Results are presented as the mean ± S.D. of three independent experiments. *, p < 0.05, compared to shCtrl cells or indicated cells. (E,F) The migration and invasion abilities of HCT116-knocked down CARMA3 cells with ectopic overexpression of Slug or vector control were performed in a Transwell assay. Results are shown as the mean ± S.D. of three independent experiments. *, p < 0.05, compared to indicated cells. Magnification: 40×. Scale bar, 100 μm. less

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Protective function of interleukin-22 in pulmonary fibrosis.

In Clin Transl Med on 1 August 2021 by Gu, P., Wang, D., et al.

Fig.2.H
Fig.2.H showing Western Blotting from the publication: Protective function of interleukin-22 in pulmonary fibrosis.
  • WB
Increased expressions of transforming growth factor (TGF)‐β1, TGF‐βR2, P‐smad2/3, and Collagen‐I in the lung of patients with idiopathic pulmonary fibrosis (IPF). (A, B) The expressions of TGF‐β1 in healthy controls (HC) and IPF patients by immunohistochemistry (IHC, ×200). (C, D) The expressions... more
Increased expressions of transforming growth factor (TGF)‐β1, TGF‐βR2, P‐smad2/3, and Collagen‐I in the lung of patients with idiopathic pulmonary fibrosis (IPF). (A, B) The expressions of TGF‐β1 in healthy controls (HC) and IPF patients by immunohistochemistry (IHC, ×200). (C, D) The expressions of TGF‐βR2 in HC and IPF patients by IHC (×200). (E) Western blot analysis in the lung tissues of HC (n = 4) and IPF patients (n = 4). (F) mRNA expression analysis of TGF‐β1 and TGF‐βR2 in HC and IPF patients by real‐time PCR in lung tissues. (G–H) Western blot analysis of protein levels of P‐smad2/3, Collagen‐I, α‐smooth muscle actin  (α‐SMA), and fibronectin (FN) in lung biopsy tissues from HC and IPF patients (HC, n = 4 and IPF patients, n = 4). (I–L) The quantification of (G–H). (Notes: phagocytes, type II AEC, bronchial mucosa epithelial cells, inflammatory cell) less

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Protective function of interleukin-22 in pulmonary fibrosis.

In Clin Transl Med on 1 August 2021 by Gu, P., Wang, D., et al.

Fig.4.N
Fig.4.N showing Western Blotting from the publication: Protective function of interleukin-22 in pulmonary fibrosis.
  • WB
Interleukin (IL)‐22 treatment ameliorated bleomycin (BLM)‐induced inflammation and collagen deposition. (A) Experimental schematic of the animal. Mice were intratracheally injected with 50 μl normal saline (NS) or BLM (5 mg/kg) to induced fibrosis at day 0. Intranasal instillation of 50 μl recomb... more
Interleukin (IL)‐22 treatment ameliorated bleomycin (BLM)‐induced inflammation and collagen deposition. (A) Experimental schematic of the animal. Mice were intratracheally injected with 50 μl normal saline (NS) or BLM (5 mg/kg) to induced fibrosis at day 0. Intranasal instillation of 50 μl recombinant IL‐22 (100 ng) was used on day 0, day 7, and day 14 after the treatment with BLM, parts of mice were sacrificed at day 7, day 14, and day 21. Each test was repeated three times. Six mice per group were used. (B1–3) The pathological changes of lung tissues on days 7, 14, and 21 in NS treated mice by HE staining (×200). (B4–6) The pathological changes of lung tissues on days 7, 14, and 21 in BLM treated mice by HE staining (×200). (B7–9) The pathological changes of lung tissues on days 7, 14, and 21 in both BLM and IL‐22 treated mice by HE staining (×200). (C1–3) The pathological changes of lung tissues at day 7, 14, and 21 days in NS treated mice by MS staining (×200). (C4–6) The pathological changes of lung tissues at day 7, 14, and 21 days in BLM treated mice by MS staining (×200). (C7–9) The pathological changes of lung tissues at day 7, 14, and 21 days in both BLM and IL‐22 treated mice by MS staining (×200). (D) Comparison of the pathological scores of alveolitis at 7 days in BLM (n = 6) and both BLM and IL‐22 treated mice (n = 6). (E) Comparison of the pathological scores of fibrosis at 21 days in BLM (n = 6) and both BLM and IL‐22 treated mice (n = 6). (F–G) The mRNA expression of transforming growth factor (TGF)‐β1 and TGF‐βR2 in lung tissues of BLM, NS, and both BLM and IL‐22 treated mice at day 21 by real‐time PCR. (H) The protein expression of TGF‐β1 and TGF‐βR2 in lung tissues of BLM, NS, both BLM and IL‐22 treated mice by WB. (I–J) The quantification of (G). (K) The protein expression of P‐smad2/3 in lung tissues of BLM, NS, and both BLM and IL‐22 treated mice by WB. (L) The quantification of (K). (N) The protein expression of α‐SMA, Collagen‐I, and fibronectin (FN) in lung tissues of BLM, NS, and both BLM and IL‐22 treated mice by WB. (M, O–P) The quantification of (N) less

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Arginine methylation of SHANK2 by PRMT7 promotes human breast cancer metastasis through activating endosomal FAK signalling.

In Elife on 26 August 2020 by Liu, Y., Li, L., et al.

Fig.7.A
Fig.7.A showing Western Blotting from the publication: Arginine methylation of SHANK2 by PRMT7 promotes human breast cancer metastasis through activating endosomal FAK signalling.
  • WB
SHANK2 R240 methylation promoted migration and invasion of breast cancer cells by activating endosomal FAK signals.(A) MCF7cells expressing the indicated Flag-tagged PRMT7. MCF7-Vector and MCF7-PRMT7 groups were treated with dynasore (80 µM) or DMSO as control. Western blot was performed with the... more
SHANK2 R240 methylation promoted migration and invasion of breast cancer cells by activating endosomal FAK signals.(A) MCF7cells expressing the indicated Flag-tagged PRMT7. MCF7-Vector and MCF7-PRMT7 groups were treated with dynasore (80 µM) or DMSO as control. Western blot was performed with the indicated EMT marker antibodies. (B and C) MCF7 cells expressing the indicated Flag-tagged PRMT7. MCF7-Vector and MCF7-PRMT7 groups were treated with dynasore (80 µM) or DMSO as control. Cell migration (B) and Invasion (C) were determined by transwell assays. (n = 3, independent experiments). Scale bars: 50 µm. Data represent the mean ± SD of three independent experiments. Scale bars,100 µm. ***p<0.001, Student’s t test. (D) MDA-MB-231 cells expressing SHANK2 shRNA with or without reconstituted expression of WT SHANK2, SHANK2 R240K or SHANK2 R240F mutant. SHANK2 R240F group was treated with dynasore (80 µM) or DMSO as control. Western blot was performed with the indicated EMT marker antibodies. (E and F) MDA-MB-231 cells expressing SHANK2 shRNA with or without reconstituted expression of WT SHANK2, SHANK2 R240K or SHANK2 R240F mutant. SHANK2 R240F group was treated with dynasore (80 µM) or DMSO as control. Cell migration (E) and Invasion (F) were determined by transwell assays. (n = 3, independent experiments). Scale bars: 100 µm. ***p<0.001, Student’s t test. (G) MDA-MB-231 cells expressing SHANK2 shRNA with or without reconstituted expression of WT SHANK2, SHANK2 R240K or SHANK2 R240F mutant. SHANK2 R240F with or without stable expression of the indicated EEA1 shRNA or a control shRNA. Western blot was performed with the indicated EMT marker antibodies. (H and I) MDA-MB-231 cells expressing SHANK2 shRNA with or without reconstituted expression of WT SHANK2, SHANK2 R240K or SHANK2 R240F mutant. SHANK2 R240F with or without stable expression of the indicated EEA1 shRNA or a control shRNA. Cell migration (H) and Invasion (I) were determined by transwell assays. (n = 3, independent experiments). Scale bars: 100 µm. ***p<0.001, Student’s t test. (J) MDA-MB-231 cells expressing SHANK2 shRNA with or without reconstituted expression of WT SHANK2, SHANK2 R240K or SHANK2 R240F mutant. SHANK2 R240F group was treated with GSK2256098 (10 µM) or DMSO as control. Western blot was performed with the indicated EMT marker antibodies. less

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Fibroblast-enriched endoplasmic reticulum protein TXNDC5 promotes pulmonary fibrosis by augmenting TGFβ signaling through TGFBR1 stabilization.

In Nat Commun on 26 August 2020 by Lee, T. H., Yeh, C. F., et al.

Fig.5.A
Fig.5.A showing Western Blotting from the publication: Fibroblast-enriched endoplasmic reticulum protein TXNDC5 promotes pulmonary fibrosis by augmenting TGFβ signaling through TGFBR1 stabilization.
  • WB
TXNDC5 is both essential and sufficient for HPF activation, proliferation, and ECM production.Protein (n = 6 biologically independent samples per group) (a) and transcript (n = 6 biologically independent samples per group) (b) expression levels of COL1A1, fibronectin, elastin, periostin, and αSMA... more
TXNDC5 is both essential and sufficient for HPF activation, proliferation, and ECM production.Protein (n = 6 biologically independent samples per group) (a) and transcript (n = 6 biologically independent samples per group) (b) expression levels of COL1A1, fibronectin, elastin, periostin, and αSMA/ACTA2 were markedly increased in control (shScr) human pulmonary fibroblasts (HPF) following TGFβ1 (10 ng/ml) treatment. TGFβ1-induced upregulation of these fibrogenic proteins/genes was significantly attenuated in HPF with TXNDC5 depletion (shTXNDC5). c TGFβ1 treatment significantly increased the cellular proliferation activity in shScr-, but not in shTXNDC5-, transduced HPF (n = 24 biologically independent samples per group). ECM proteins (COL1A1 and fibronectin), markers for fibroblast activation (periostin and αSMA) (n = 9 biologically independent samples per group) (d) and fibroblast proliferation activity (n = 9 biologically independent samples per group) (e) were markedly increased in HPF transduced with TXNDC5 (TXNDC5 OE), compared with empty, expression vector (Data are presented as mean ± SEM, P value determined using two-tailed Mann–Whitney U test. Source data are provided as a Source Data file. n.s. non-significant, ctrl control, KD knockdown, OE overexpress). less

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Arginine methylation of SHANK2 by PRMT7 promotes human breast cancer metastasis through activating endosomal FAK signalling.

In Elife on 26 August 2020 by Liu, Y., Li, L., et al.

Fig.7.D
Fig.7.D showing Western Blotting from the publication: Arginine methylation of SHANK2 by PRMT7 promotes human breast cancer metastasis through activating endosomal FAK signalling.
  • WB
SHANK2 R240 methylation promoted migration and invasion of breast cancer cells by activating endosomal FAK signals.(A) MCF7cells expressing the indicated Flag-tagged PRMT7. MCF7-Vector and MCF7-PRMT7 groups were treated with dynasore (80 µM) or DMSO as control. Western blot was performed with the... more
SHANK2 R240 methylation promoted migration and invasion of breast cancer cells by activating endosomal FAK signals.(A) MCF7cells expressing the indicated Flag-tagged PRMT7. MCF7-Vector and MCF7-PRMT7 groups were treated with dynasore (80 µM) or DMSO as control. Western blot was performed with the indicated EMT marker antibodies. (B and C) MCF7 cells expressing the indicated Flag-tagged PRMT7. MCF7-Vector and MCF7-PRMT7 groups were treated with dynasore (80 µM) or DMSO as control. Cell migration (B) and Invasion (C) were determined by transwell assays. (n = 3, independent experiments). Scale bars: 50 µm. Data represent the mean ± SD of three independent experiments. Scale bars,100 µm. ***p<0.001, Student’s t test. (D) MDA-MB-231 cells expressing SHANK2 shRNA with or without reconstituted expression of WT SHANK2, SHANK2 R240K or SHANK2 R240F mutant. SHANK2 R240F group was treated with dynasore (80 µM) or DMSO as control. Western blot was performed with the indicated EMT marker antibodies. (E and F) MDA-MB-231 cells expressing SHANK2 shRNA with or without reconstituted expression of WT SHANK2, SHANK2 R240K or SHANK2 R240F mutant. SHANK2 R240F group was treated with dynasore (80 µM) or DMSO as control. Cell migration (E) and Invasion (F) were determined by transwell assays. (n = 3, independent experiments). Scale bars: 100 µm. ***p<0.001, Student’s t test. (G) MDA-MB-231 cells expressing SHANK2 shRNA with or without reconstituted expression of WT SHANK2, SHANK2 R240K or SHANK2 R240F mutant. SHANK2 R240F with or without stable expression of the indicated EEA1 shRNA or a control shRNA. Western blot was performed with the indicated EMT marker antibodies. (H and I) MDA-MB-231 cells expressing SHANK2 shRNA with or without reconstituted expression of WT SHANK2, SHANK2 R240K or SHANK2 R240F mutant. SHANK2 R240F with or without stable expression of the indicated EEA1 shRNA or a control shRNA. Cell migration (H) and Invasion (I) were determined by transwell assays. (n = 3, independent experiments). Scale bars: 100 µm. ***p<0.001, Student’s t test. (J) MDA-MB-231 cells expressing SHANK2 shRNA with or without reconstituted expression of WT SHANK2, SHANK2 R240K or SHANK2 R240F mutant. SHANK2 R240F group was treated with GSK2256098 (10 µM) or DMSO as control. Western blot was performed with the indicated EMT marker antibodies. less

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Arginine methylation of SHANK2 by PRMT7 promotes human breast cancer metastasis through activating endosomal FAK signalling.

In Elife on 26 August 2020 by Liu, Y., Li, L., et al.

Fig.7.G
Fig.7.G showing Western Blotting from the publication: Arginine methylation of SHANK2 by PRMT7 promotes human breast cancer metastasis through activating endosomal FAK signalling.
  • WB
SHANK2 R240 methylation promoted migration and invasion of breast cancer cells by activating endosomal FAK signals.(A) MCF7cells expressing the indicated Flag-tagged PRMT7. MCF7-Vector and MCF7-PRMT7 groups were treated with dynasore (80 µM) or DMSO as control. Western blot was performed with the... more
SHANK2 R240 methylation promoted migration and invasion of breast cancer cells by activating endosomal FAK signals.(A) MCF7cells expressing the indicated Flag-tagged PRMT7. MCF7-Vector and MCF7-PRMT7 groups were treated with dynasore (80 µM) or DMSO as control. Western blot was performed with the indicated EMT marker antibodies. (B and C) MCF7 cells expressing the indicated Flag-tagged PRMT7. MCF7-Vector and MCF7-PRMT7 groups were treated with dynasore (80 µM) or DMSO as control. Cell migration (B) and Invasion (C) were determined by transwell assays. (n = 3, independent experiments). Scale bars: 50 µm. Data represent the mean ± SD of three independent experiments. Scale bars,100 µm. ***p<0.001, Student’s t test. (D) MDA-MB-231 cells expressing SHANK2 shRNA with or without reconstituted expression of WT SHANK2, SHANK2 R240K or SHANK2 R240F mutant. SHANK2 R240F group was treated with dynasore (80 µM) or DMSO as control. Western blot was performed with the indicated EMT marker antibodies. (E and F) MDA-MB-231 cells expressing SHANK2 shRNA with or without reconstituted expression of WT SHANK2, SHANK2 R240K or SHANK2 R240F mutant. SHANK2 R240F group was treated with dynasore (80 µM) or DMSO as control. Cell migration (E) and Invasion (F) were determined by transwell assays. (n = 3, independent experiments). Scale bars: 100 µm. ***p<0.001, Student’s t test. (G) MDA-MB-231 cells expressing SHANK2 shRNA with or without reconstituted expression of WT SHANK2, SHANK2 R240K or SHANK2 R240F mutant. SHANK2 R240F with or without stable expression of the indicated EEA1 shRNA or a control shRNA. Western blot was performed with the indicated EMT marker antibodies. (H and I) MDA-MB-231 cells expressing SHANK2 shRNA with or without reconstituted expression of WT SHANK2, SHANK2 R240K or SHANK2 R240F mutant. SHANK2 R240F with or without stable expression of the indicated EEA1 shRNA or a control shRNA. Cell migration (H) and Invasion (I) were determined by transwell assays. (n = 3, independent experiments). Scale bars: 100 µm. ***p<0.001, Student’s t test. (J) MDA-MB-231 cells expressing SHANK2 shRNA with or without reconstituted expression of WT SHANK2, SHANK2 R240K or SHANK2 R240F mutant. SHANK2 R240F group was treated with GSK2256098 (10 µM) or DMSO as control. Western blot was performed with the indicated EMT marker antibodies. less

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Arginine methylation of SHANK2 by PRMT7 promotes human breast cancer metastasis through activating endosomal FAK signalling.

In Elife on 26 August 2020 by Liu, Y., Li, L., et al.

Fig.7.J
Fig.7.J showing Western Blotting from the publication: Arginine methylation of SHANK2 by PRMT7 promotes human breast cancer metastasis through activating endosomal FAK signalling.
  • WB
SHANK2 R240 methylation promoted migration and invasion of breast cancer cells by activating endosomal FAK signals.(A) MCF7cells expressing the indicated Flag-tagged PRMT7. MCF7-Vector and MCF7-PRMT7 groups were treated with dynasore (80 µM) or DMSO as control. Western blot was performed with the... more
SHANK2 R240 methylation promoted migration and invasion of breast cancer cells by activating endosomal FAK signals.(A) MCF7cells expressing the indicated Flag-tagged PRMT7. MCF7-Vector and MCF7-PRMT7 groups were treated with dynasore (80 µM) or DMSO as control. Western blot was performed with the indicated EMT marker antibodies. (B and C) MCF7 cells expressing the indicated Flag-tagged PRMT7. MCF7-Vector and MCF7-PRMT7 groups were treated with dynasore (80 µM) or DMSO as control. Cell migration (B) and Invasion (C) were determined by transwell assays. (n = 3, independent experiments). Scale bars: 50 µm. Data represent the mean ± SD of three independent experiments. Scale bars,100 µm. ***p<0.001, Student’s t test. (D) MDA-MB-231 cells expressing SHANK2 shRNA with or without reconstituted expression of WT SHANK2, SHANK2 R240K or SHANK2 R240F mutant. SHANK2 R240F group was treated with dynasore (80 µM) or DMSO as control. Western blot was performed with the indicated EMT marker antibodies. (E and F) MDA-MB-231 cells expressing SHANK2 shRNA with or without reconstituted expression of WT SHANK2, SHANK2 R240K or SHANK2 R240F mutant. SHANK2 R240F group was treated with dynasore (80 µM) or DMSO as control. Cell migration (E) and Invasion (F) were determined by transwell assays. (n = 3, independent experiments). Scale bars: 100 µm. ***p<0.001, Student’s t test. (G) MDA-MB-231 cells expressing SHANK2 shRNA with or without reconstituted expression of WT SHANK2, SHANK2 R240K or SHANK2 R240F mutant. SHANK2 R240F with or without stable expression of the indicated EEA1 shRNA or a control shRNA. Western blot was performed with the indicated EMT marker antibodies. (H and I) MDA-MB-231 cells expressing SHANK2 shRNA with or without reconstituted expression of WT SHANK2, SHANK2 R240K or SHANK2 R240F mutant. SHANK2 R240F with or without stable expression of the indicated EEA1 shRNA or a control shRNA. Cell migration (H) and Invasion (I) were determined by transwell assays. (n = 3, independent experiments). Scale bars: 100 µm. ***p<0.001, Student’s t test. (J) MDA-MB-231 cells expressing SHANK2 shRNA with or without reconstituted expression of WT SHANK2, SHANK2 R240K or SHANK2 R240F mutant. SHANK2 R240F group was treated with GSK2256098 (10 µM) or DMSO as control. Western blot was performed with the indicated EMT marker antibodies. less

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Arginine methylation of SHANK2 by PRMT7 promotes human breast cancer metastasis through activating endosomal FAK signalling.

In Elife on 26 August 2020 by Liu, Y., Li, L., et al.

Fig.3.A
Fig.3.A showing Western Blotting from the publication: Arginine methylation of SHANK2 by PRMT7 promotes human breast cancer metastasis through activating endosomal FAK signalling.
  • WB
SHANK2 was involved in breast cancer cells EMT.(A) BT549 cells with or without expressing SHANK2 shRNA and with or without reconstituted expression of SHANK2 WT or SHANK2 R240K mutant. Western blot was performed with the indicated EMT marker antibodies. (B) MDA-MB-231 with or without stable expre... more
SHANK2 was involved in breast cancer cells EMT.(A) BT549 cells with or without expressing SHANK2 shRNA and with or without reconstituted expression of SHANK2 WT or SHANK2 R240K mutant. Western blot was performed with the indicated EMT marker antibodies. (B) MDA-MB-231 with or without stable expression of the indicated SHANK2 shRNA or a control shRNA. Western blot was performed with the indicated EMT marker antibodies. (C) MCF10A cells expressing the indicated HA-tagged SHANK2. Western blot was performed with the indicated EMT marker antibodies. (D) Stem-like CD44high/CD24low profile in MDA-MB-231 cells transfected with stable expression of the indicated SHANK2 shRNA or a control shRNA, and MCF10A cells expressing the indicated HA-tagged SHANK2. less

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Low levels of AMPK promote epithelial-mesenchymal transition in lung cancer primarily through HDAC4- and HDAC5-mediated metabolic reprogramming.

In J Cell Mol Med on 1 July 2020 by Feng, S., Zhang, L., et al.

Fig.4.E
Fig.4.E showing Western Blotting from the publication: Low levels of AMPK promote epithelial-mesenchymal transition in lung cancer primarily through HDAC4- and HDAC5-mediated metabolic reprogramming.
  • WB
Low AMPK induces cell EMT by promoting glycolysis in HBE cells. A, High‐performance liquid chromatography (HPLC) analysis for ATP in HBE cells, **P < .01 vs the vector group; NS, P > .05 vs the control group, n = 3. B, Relative glycolysis rates in HBE cells, as judged by Seahorse analyses, **P < ... more
Low AMPK induces cell EMT by promoting glycolysis in HBE cells. A, High‐performance liquid chromatography (HPLC) analysis for ATP in HBE cells, **P < .01 vs the vector group; NS, P > .05 vs the control group, n = 3. B, Relative glycolysis rates in HBE cells, as judged by Seahorse analyses, **P < .01 vs the vector group; NS, P > .05 vs the control group, n = 3. C, D, Metabolic analyses were done with the Seahorse XFe96 Analyzer, which measures the ECAR and OCR of live HBE cells, ***P < .001 vs the vector group; NS, P > .05 vs the control group, n = 3. E, F, Western blot determination of EMT‐related markers in response to 2‐DG, n = 3. G‐L, Relative mRNA expression levels of EMT markers and inducer in HBE cells, *P < .05, **P < .01, ***P < .001, NS, P > .05 vs the indicated group, n = 3 less

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Low levels of AMPK promote epithelial-mesenchymal transition in lung cancer primarily through HDAC4- and HDAC5-mediated metabolic reprogramming.

In J Cell Mol Med on 1 July 2020 by Feng, S., Zhang, L., et al.

Fig.4.F
Fig.4.F showing Western Blotting from the publication: Low levels of AMPK promote epithelial-mesenchymal transition in lung cancer primarily through HDAC4- and HDAC5-mediated metabolic reprogramming.
  • WB
Low AMPK induces cell EMT by promoting glycolysis in HBE cells. A, High‐performance liquid chromatography (HPLC) analysis for ATP in HBE cells, **P < .01 vs the vector group; NS, P > .05 vs the control group, n = 3. B, Relative glycolysis rates in HBE cells, as judged by Seahorse analyses, **P < ... more
Low AMPK induces cell EMT by promoting glycolysis in HBE cells. A, High‐performance liquid chromatography (HPLC) analysis for ATP in HBE cells, **P < .01 vs the vector group; NS, P > .05 vs the control group, n = 3. B, Relative glycolysis rates in HBE cells, as judged by Seahorse analyses, **P < .01 vs the vector group; NS, P > .05 vs the control group, n = 3. C, D, Metabolic analyses were done with the Seahorse XFe96 Analyzer, which measures the ECAR and OCR of live HBE cells, ***P < .001 vs the vector group; NS, P > .05 vs the control group, n = 3. E, F, Western blot determination of EMT‐related markers in response to 2‐DG, n = 3. G‐L, Relative mRNA expression levels of EMT markers and inducer in HBE cells, *P < .05, **P < .01, ***P < .001, NS, P > .05 vs the indicated group, n = 3 less

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Low levels of AMPK promote epithelial-mesenchymal transition in lung cancer primarily through HDAC4- and HDAC5-mediated metabolic reprogramming.

In J Cell Mol Med on 1 July 2020 by Feng, S., Zhang, L., et al.

Fig.5.H
Fig.5.H showing Western Blotting from the publication: Low levels of AMPK promote epithelial-mesenchymal transition in lung cancer primarily through HDAC4- and HDAC5-mediated metabolic reprogramming.
  • WB
Low AMPK promotes glycolysis by up‐regulating the expression of HK2 in HBE cells. A, B, Relative mRNA and protein expression levels of HK1, HK2, LDHA, G6PI,TPI, MCT‐1 and PFK‐1 in HBE cells, *P < .05, ***P < .001, vs the Vector group, n = 3. C, D, Relative mRNA and protein expression levels of HK... more
Low AMPK promotes glycolysis by up‐regulating the expression of HK2 in HBE cells. A, B, Relative mRNA and protein expression levels of HK1, HK2, LDHA, G6PI,TPI, MCT‐1 and PFK‐1 in HBE cells, *P < .05, ***P < .001, vs the Vector group, n = 3. C, D, Relative mRNA and protein expression levels of HK2, AMPK and p‐AMPK in HBE cells. E‐G, Relative glycolysis rates, migration, invasion in response to siHK2 treatment, *P < .05, **P < .01, ***P < .001 vs the respective vector group in HBE cells, n = 3. H, Western blot determination of EMT‐related markers protein expression in response to siHK2 treatment in sgAMPK HBE cells, *P < .05, **P < .01, ***P < .001 vs the vector group, n = 3. I‐K, Relative glycolysis rates, migration, invasion in response to siHK2 treatment, *P < .05, **P < .01, ***P < .001 vs the respective vector group in sgAMPK HBE cells, n = 3 less

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Inhibition of TGF-β signalling in combination with nal-IRI plus 5-Fluorouracil/Leucovorin suppresses invasion and prolongs survival in pancreatic tumour mouse models.

In Sci Rep on 19 February 2020 by Hong, E., Park, S., et al.

Fig.5.D
Fig.5.D showing Western Blotting in a Homo sapiens (Human) sample from the publication: Inhibition of TGF-β signalling in combination with nal-IRI plus 5-Fluorouracil/Leucovorin suppresses invasion and prolongs survival in pancreatic tumour mouse models.
  • WB
  • Homo sapiens (Human)
Ectopic expression of CCDC80 reducing migration, colony formation, and EMT in pancreatic cancer cells. (A) Transwell migration assay measuring migration abilities of PANC-1 and Panc02 cells expressing LPCX or CCDC80. (B) Representative images of colonies stained with methylene blue in PANC-1 and ... more
Ectopic expression of CCDC80 reducing migration, colony formation, and EMT in pancreatic cancer cells. (A) Transwell migration assay measuring migration abilities of PANC-1 and Panc02 cells expressing LPCX or CCDC80. (B) Representative images of colonies stained with methylene blue in PANC-1 and Panc02 cells stably expressing CCDC80. (C) qRT-PCR results showing down-regulation of EMT marker expression in PANC-1 and Panc02 cells by ectopic expression of CCDC80. All the data is represented as the mean of three repeated values. ***P < 0.0005, **P < 0.005, and *P < 0.05 compared to the control. (D) Western blot analysis showing reduction of EMT markers in PANC-1 and Panc02 with stably expressing CCDC80. The blots are cropped, and the original blots are presented in Supplementary Fig. 8. Note that overexpression of CCDC80 decreases EMT marker expression in pancreatic cancer cells. less

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Inhibition of TGF-β signalling in combination with nal-IRI plus 5-Fluorouracil/Leucovorin suppresses invasion and prolongs survival in pancreatic tumour mouse models.

In Sci Rep on 19 February 2020 by Hong, E., Park, S., et al.

Fig.2.C
Fig.2.C showing Western Blotting in a Homo sapiens (Human) sample from the publication: Inhibition of TGF-β signalling in combination with nal-IRI plus 5-Fluorouracil/Leucovorin suppresses invasion and prolongs survival in pancreatic tumour mouse models.
  • WB
  • Homo sapiens (Human)
Synergistic effect of vactosertib with nal-IRI/5-FU on migration, invasion, and EMT of pancreatic cancer cells. (A) Transwell cell migration assay. Cells were treated with the indicated reagent(s) and incubated for 48 hours before placed in a migration chamber. 1 × 105 of PANC-1 (left) or 5 × 104... more
Synergistic effect of vactosertib with nal-IRI/5-FU on migration, invasion, and EMT of pancreatic cancer cells. (A) Transwell cell migration assay. Cells were treated with the indicated reagent(s) and incubated for 48 hours before placed in a migration chamber. 1 × 105 of PANC-1 (left) or 5 × 104 of Panc02 (right) cells were then seeded in migration chambers for 48 hours. The relative number of migrated cells in each group was counted. (B) Cell invasion assay. Cells were treated with the indicated reagent(s) and incubated for 48 hours before placed in an invasion chamber. 2 × 105 PANC-1 (left) or 1 × 105 Panc02 (right) cells were then seeded in invasion chambers for 48 hours. The number of invaded cells in each group was counted. Note that the combination of vactosertib with nal-IRI/5-FU inhibits migration and invasion in PANC-1 and Panc02 pancreatic cancer cells. (A,B) The values for migrated/invaded number of cells represent the mean ± SD of triplicate data. ***P < 0.0005, **P < 0.005, and *P < 0.05 compared to the control group; ##P < 0.005 and #P < 0.05 compared to the nal-IRI-treated group. (C) Western blot analysis for EMT markers. Vactosertib was treated for 2 hours prior to incubation with nal-IRI/5-FU in the presence of 5 ng/ml TGF-β for 48 hours. The blots are cropped and the full-length images are presented in Supplementary Fig. 8. Note that vactosertib and its combination with nal-IRI/5-FU restore the TGF-β-mediated reduction of E-cadherin and the increase of mesenchymal markers. less

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Quercetin Inhibits Cell Survival and Metastatic Ability via the EMT-mediated Pathway in Oral Squamous Cell Carcinoma.

In Molecules on 10 February 2020 by Kim, S. R., Lee, E. Y., et al.

Fig.3.A
Fig.3.A showing Western Blotting from the publication: Quercetin Inhibits Cell Survival and Metastatic Ability via the EMT-mediated Pathway in Oral Squamous Cell Carcinoma.
  • WB
Quercetin is shown to induce regulation of epithelial-mesenchymal transition (EMT) and matrix metalloproteinase (MMP). (A) Western blotting was conducted to examine the changes in the EMT inducers. The results showed that the epithelial markers (E-cadherin and claudin-1) were upregulated, and the... more
Quercetin is shown to induce regulation of epithelial-mesenchymal transition (EMT) and matrix metalloproteinase (MMP). (A) Western blotting was conducted to examine the changes in the EMT inducers. The results showed that the epithelial markers (E-cadherin and claudin-1) were upregulated, and the mesenchymal markers (fibronectin, vimentin, and alpha-smooth muscle actin (α-SMA)) were downregulated upon treatment with quercetin. (B) Quantitation of A. The band intensities of each target protein were measured using an image analyzer and presented as relative ratio. (C) Gelatin zymography shows the MMP-2 and MMP-9 activities in oral cancer cell lines (OSC20, SAS, and HN22 cells) upon quercetin treatment. Data are the means ± SEM. * p < 0.05, and ** p < 0.01 vs. the corresponding control (quercetin 0 μM). less

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