Regulation of aquaporin-2 (Aqp2) gene is essential for body water homeostasis. This study investigated how TAZ (a transcriptional coactivator with PDZ-binding motif, Wwtr1) controls vasopressin-driven AQP2 expression. AQP2 expression was studied using collecting duct-specific TAZ-knockout (TAZf/f; HoxB7Cre) mice and siRNA-mediated knockdown of TAZ in vasopressin-responsive mpkCCDc11 cells. Downstream factors of TAZ were identified using transcriptomics and bioinformatics. The TAZf/f; HoxB7Cre mice demonstrated polyuria and a significant decrease in AQP2 abundance in the kidney cortex and the outer medulla. dDAVP treatment (10-9 M, 24 h) on mpkCCDc11 cells significantly increased AQP2 mRNA and protein levels. However, siRNA-mediated TAZ knockdown (TAZ-KD) markedly attenuated these effects without affecting cAMP levels. Immunocytochemical analysis revealed a substantial decrease in AQP2 immunolabeling intensity in TAZ-KD cells following dDAVP stimulation. RNA sequencing analysis identified 1370 and 1985 differentially expressed genes in TAZ-KD cells under basal conditions and after dDAVP treatment, respectively. Among 17 previously identified transcription factor (TF) candidates, seven (Nr4a1, Cebpb, Mef2d, Elf3, Klf5, Junb, Stat3) were significantly upregulated by dDAVP in either control or TAZ-KD conditions. Among them, RT-qPCR analysis identified Nr4a1 as a TAZ-dependent TF, and immunoblotting revealed reduced NR4A1 protein levels in TAZ-KD cells upon dDAVP stimulation. This finding suggests its role as a TAZ-regulated target in dDAVP response pathway. Accordingly, Nr4a1-KD reduced the dDAVP-induced upregulation of Aqp2 mRNA and protein. KEGG pathway enrichment analysis revealed that HIF-1 signaling and glycolysis as central pathways affected by TAZ. TAZ-NR4A1 axis acts as a novel transcriptional regulatory mechanism in controlling vasopressin-mediated AQP2 expression.
© 2025 The Author(s). The FASEB Journal published by Wiley Periodicals LLC on behalf of Federation of American Societies for Experimental Biology.
Product Citations: 105
Vasopressin-Sensitive Aqp2 Regulation Mediated by the TAZ-NR4A1 Axis in Renal Collecting Duct Cells.
In The FASEB Journal on 15 July 2025 by Choi, H. S., Jang, H. J., et al.
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Endocrinology and Physiology
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C1orf50 Drives Malignant Melanoma Progression Through the Regulation of Stemness.
In Cancer Genomics Proteomics on 26 June 2025 by Otani, Y., Maekawa, M., et al.
Recent advancements in omics analysis have significantly enhanced our understanding of the molecular pathology of malignant melanoma, leading to the development of novel therapeutic strategies that target specific vulnerabilities within the disease. Despite these improvements, the factors contributing to the poor prognosis of patients with malignant melanoma remain incompletely understood. The aim of this study was to investigate the role of C1orf50 (Chromosome 1 open reading frame 50), a gene previously of unknown function, as a prognostic biomarker in melanoma.
We performed comprehensive transcriptome data analysis and subsequent functional validation of the human Skin Cutaneous Melanoma project from The Cancer Genome Atlas (TCGA).
Elevated expression levels of C1orf50 correlated with worse survival outcomes. Mechanistically, we revealed that C1orf50 plays a significant role in the regulation of cell cycle processes and cancer cell stemness, providing a potential avenue for novel therapeutic interventions in melanoma.
This study is the first to identify C1orf50 as a prognostic biomarker in melanoma. The clinical relevance of our results sheds light on the importance of further investigation into the biological mechanisms underpinning C1orf50's impact on melanoma progression and patient prognosis.
Copyright © 2025, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.
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Cancer Research
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In Development Reproduction on 1 June 2025 by Hwang, S., Kim, B., et al.
Transcriptional coactivator with PDZ-binding motif (TAZ) functions as a transcriptional coactivator, which shuttles between the cytoplasm and the nucleus under the Hippo signaling. It is known to be involved in promoting cell proliferation, organ overgrowth, survival to stress, and dedifferentiation by interacting with TEAD transcription factors (TEADs). However, the regulation of TAZ by intrauterine hormones has not yet been investigated. In this study, we investigated TAZ expression during the estrous cycle in the normal mouse uterus and the effect of estrogen and progesterone on TAZ expression in the ovariectomized (OVX) mouse uterus. TAZ expression levels did not show a statistically significant change in the uterus during the estrous cycle. However, immunofluorescence revealed that TAZ nuclear localization significantly increased at the estrus stage. In the OVX mouse uterus, the expression levels of TAZ mRNA and protein dramatically increased in a time-dependent manner after estrogen treatment. Also, immunofluorescence showed that the nuclear TAZ expression increased at 6 h and 12 h after estrogen treatment compared to the oil treated OVX mouse uterus (0 h). Finally, pretreatment of an estrogen receptor (ER) antagonist ICI 182,780 efficiently reduced estrogen-induced TAZ expression. However, progesterone did not significantly affect the expression of TAZ in both mRNA and protein levels. In conclusion, TAZ expression is regulated and activated by estrogen through nuclear estrogen receptors, ERα, and ERβ in the uterine environment.
© Copyright 2025 The Korean Society of Developmental Biology.
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Biochemistry and Molecular biology
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Endocrinology and Physiology
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In Cell Death and Differentiation on 1 May 2025 by Zhu, Z., Ding, R., et al.
Hyperactivation of the YAP/TEAD transcriptional complex in cancers facilitates the development of an immunosuppressive tumor microenvironment. Herein, we observed that the transcription factor SP1 physically interacts with and stabilizes the YAP/TEAD complex at regulatory genomic loci in colorectal cancer (CRC). In response to serum stimulation, PKCζ (protein kinase C ζ) was found to phosphorylate SP1 and enhance its interaction with TEAD4. As a result, SP1 enhanced the transcriptional activity of YAP/TEAD and coregulated the expression of a group of YAP/TEAD target genes. The immune checkpoint V-domain Ig suppressor of T-cell activation (VISTA) was identified as a direct target of the SP1-YAP/TEAD4 complex and found to be widely expressed in CRC cells. Importantly, YAP-induced VISTA upregulation in human CRC cells was found to strongly suppress the antitumor function of CD8+ T cells. Consistently, elevated VISTA expression was found to be correlated with hyperactivation of the SP1-YAP/TEAD axis and associated with poor prognosis of CRC patients. In addition, we found by serendipity that enzymatic deglycosylation significantly improved the anti-VISTA antibody signal intensity, resulting in more accurate detection of VISTA in clinical tumor samples. Overall, our study identified SP1 as a positive modulator of YAP/TEAD for the transcriptional regulation of VISTA and developed a protein deglycosylation strategy to better detect VISTA expression in clinical samples. These findings revealed a new tumor cell-intrinsic mechanism of YAP/TAZ-mediated cancer immune evasion.
© 2025. The Author(s).
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PLA
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Cancer Research
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Cell Biology
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In IScience on 18 April 2025 by Kofler, M., Venugopal, S., et al.
Multiple mechanisms were proposed to mediate the nuclear import of TAZ/YAP, transcriptional co-activators regulating organ growth and regeneration. Our earlier observations showed that TAZ/YAP harbor a C-terminal, unconventional nuclear localization signal (NLS). Here, we show that this sequence, necessary and sufficient for basal, ATP-independent nuclear import, contains an indispensable central methionine flanked by negatively charged residues. Based on these features, we define the M-motif and propose that it is a new class of NLS, also present and import-competent in other cellular (STAT1 and cyclin B1) and viral (ORF6 of SARS-CoV2, VSV-M) proteins. Accordingly, ORF6 SARS-Cov2 competitively inhibits TAZ/YAP uptake, while TAZ abrogates STAT1 import. Similar to viral M-motif proteins, TAZ binds RAE1 and inhibits classic nuclear protein import, including that of antiviral factors (IRF3 and NF-κB). However, RAE1 is dispensable for TAZ import itself. Thus, the TAZ/YAP NLS has a dual function: it mediates unconventional nuclear import and inhibits classic import, contributing to the suppression of antiviral responses.
© 2025 The Authors.
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Immunology and Microbiology
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In Nat Commun on 10 July 2024 by Patterson, M. R., Cogan, J. A., et al.
Fig.1.D
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TAZ is upregulated in HPV18+ cervical cancer.A Genomic alterations of WWTR1 across human cancers determined by cBioportal analysis of TCGA data (LUSC Lung squamous cell carcinoma (n = 487), USC Uterine serous carcinoma (n = 56), ESCA Eosophageal carcinoma (n = 186), OV Ovarian serous cystadenocar...
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TAZ is upregulated in HPV18+ cervical cancer.A Genomic alterations of WWTR1 across human cancers determined by cBioportal analysis of TCGA data (LUSC Lung squamous cell carcinoma (n = 487), USC Uterine serous carcinoma (n = 56), ESCA Eosophageal carcinoma (n = 186), OV Ovarian serous cystadenocarcinoma (n = 579), HNSC Head and Neck squamous cell carcinoma (n = 517), CESC Cervical squamous cell carcinoma (n = 295), BLCA Bladder Urothelial carcinoma (n = 408), PRAD Prostate adenocarcinoma (n = 491), UCEC Uterine corpus endometrial carcinoma (n = 487). B Scatter dot plot analysis of WWTR1 mRNA expression against WWTR1 copy number alterations in cervical cancer determined from TCGA data (n = 278). The correlation coefficient (r) was calculated using Pearson correlation analysis. C Scatter dot plot analysis of WWTR1 mRNA expression against TAZ protein expression in cervical cancer determined from TCGA data (n = 152). The correlation coefficient (r) was calculated using Pearson correlation analysis. D Representative western blot of lysate from HPV-, HPV16+ and HPV18+ cell lines. Lysates were probed for YAP, TAZ and the loading control GAPDH (n = 3). E RT-qPCR analysis of WWTR1 or YAP1 expression in HPV-, HPV16+ or HPV18+ cell lines (n = 3). U6 transcript levels were used as a loading control. F RT-qPCR analysis of WWTR1 expression in negative, HPV16 or HPV18+ patient cervix liquid cytology samples from different CIN grades (n = 4 from each grade). U6 was used as a loading control. Maxima and minima whiskers represent the highest and lowest values, respectively. Bounds of boxes are the 75th and 25th percentiles and the centre represents the median. G Kaplan–Meier curves showing progression free survival in HPV18+ cervical cancer stratified by high or low WWTR1 expression. Survival was compared using the log-rank test. H Kaplan–Meier curves showing progression free survival in non-HPV18+ cervical cancer stratified by high or low WWTR1 expression. Survival was compared using the log-rank test. Error bars represent the mean +/- standard deviation of a minimum of three biological repeats when relevant. *P < 0.05, **P < 0.01, ***P < 0.005 (two-tailed, unpaired Student’s t-test). Source data are provided as a Source Data file.
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Collected and cropped from Nature Communications by CiteAb, provided under a CC-BY license
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In Nat Commun on 10 July 2024 by Patterson, M. R., Cogan, J. A., et al.
Fig.2.G
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HPV18 E7 upregulates WWTR1 transcription.A Representative western blots of HeLa cells after transfection with HPV18 E6/E7 targeting siRNA. Cell lysates were probed for TAZ, YAP, HPV18 E6 and HPV18 E7 expression. GAPDH was used as a loading control (n = 4). B RT-qPCR analysis of WWTR1 expression i...
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HPV18 E7 upregulates WWTR1 transcription.A Representative western blots of HeLa cells after transfection with HPV18 E6/E7 targeting siRNA. Cell lysates were probed for TAZ, YAP, HPV18 E6 and HPV18 E7 expression. GAPDH was used as a loading control (n = 4). B RT-qPCR analysis of WWTR1 expression in HeLa cells after transfection with HPV18 E6/E7 targeting siRNA (n = 3). U6 was used as a loading control. CWWTR1-promoter luciferase assay in HeLa cells after transfection with HPV18 E6/E7 targeting siRNA (n = 3). D Representative western blot of C33A cell lysates stably expressing either HA-HPV18 E6 or HA-HPV18 E7 probed for TAZ, YAP, HPV18 E6, HPV18 E7 and the loading control GAPDH (n = 3). E RT-qPCR analysis of WWTR1 expression in C33A cells stably expressing either HA-HPV18 E6 or HA-HPV18 E7 (n = 3). U6 was used as a loading control. FWWTR1-promoter luciferase assay in C33A cells stably expressing either HA-HPV18 E6 or HA-HPV18 E7 (n = 3). G Representative western blot of C33A cell lysate stably expressing either HA-HPV18 E6 or HA-HPV18 E7 probed for TAZ, YAP, HPV16 E7, HPV18 E7 and GAPDH (n = 3). H RT-qPCR analysis of WWTR1 expression in C33A cells stably expressing either HA-HPV16 E7 or HA-HPV18 E7 (n = 4). U6 was used as a loading control. IWWTR1-promoter luciferase assay in C33A cells stably expressing either HA-HPV16 E7 or HA-HPV18 E7 (n = 3). Error bars represent the mean +/- standard deviation of a minimum of three biological repeats. *P < 0.05, **P < 0.01, ***P < 0.005 (two-tailed, unpaired Student’s t-test). Source data are provided as a Source Data file.
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Collected and cropped from Nature Communications by CiteAb, provided under a CC-BY license
Image 1 of 32
In Nat Commun on 10 July 2024 by Patterson, M. R., Cogan, J. A., et al.
Fig.2.A
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HPV18 E7 upregulates WWTR1 transcription.A Representative western blots of HeLa cells after transfection with HPV18 E6/E7 targeting siRNA. Cell lysates were probed for TAZ, YAP, HPV18 E6 and HPV18 E7 expression. GAPDH was used as a loading control (n = 4). B RT-qPCR analysis of WWTR1 expression i...
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HPV18 E7 upregulates WWTR1 transcription.A Representative western blots of HeLa cells after transfection with HPV18 E6/E7 targeting siRNA. Cell lysates were probed for TAZ, YAP, HPV18 E6 and HPV18 E7 expression. GAPDH was used as a loading control (n = 4). B RT-qPCR analysis of WWTR1 expression in HeLa cells after transfection with HPV18 E6/E7 targeting siRNA (n = 3). U6 was used as a loading control. CWWTR1-promoter luciferase assay in HeLa cells after transfection with HPV18 E6/E7 targeting siRNA (n = 3). D Representative western blot of C33A cell lysates stably expressing either HA-HPV18 E6 or HA-HPV18 E7 probed for TAZ, YAP, HPV18 E6, HPV18 E7 and the loading control GAPDH (n = 3). E RT-qPCR analysis of WWTR1 expression in C33A cells stably expressing either HA-HPV18 E6 or HA-HPV18 E7 (n = 3). U6 was used as a loading control. FWWTR1-promoter luciferase assay in C33A cells stably expressing either HA-HPV18 E6 or HA-HPV18 E7 (n = 3). G Representative western blot of C33A cell lysate stably expressing either HA-HPV18 E6 or HA-HPV18 E7 probed for TAZ, YAP, HPV16 E7, HPV18 E7 and GAPDH (n = 3). H RT-qPCR analysis of WWTR1 expression in C33A cells stably expressing either HA-HPV16 E7 or HA-HPV18 E7 (n = 4). U6 was used as a loading control. IWWTR1-promoter luciferase assay in C33A cells stably expressing either HA-HPV16 E7 or HA-HPV18 E7 (n = 3). Error bars represent the mean +/- standard deviation of a minimum of three biological repeats. *P < 0.05, **P < 0.01, ***P < 0.005 (two-tailed, unpaired Student’s t-test). Source data are provided as a Source Data file.
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Collected and cropped from Nature Communications by CiteAb, provided under a CC-BY license
Image 1 of 32
In Nat Commun on 10 July 2024 by Patterson, M. R., Cogan, J. A., et al.
Fig.2.D
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Homo sapiens (Human)
HPV18 E7 upregulates WWTR1 transcription.A Representative western blots of HeLa cells after transfection with HPV18 E6/E7 targeting siRNA. Cell lysates were probed for TAZ, YAP, HPV18 E6 and HPV18 E7 expression. GAPDH was used as a loading control (n = 4). B RT-qPCR analysis of WWTR1 expression i...
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HPV18 E7 upregulates WWTR1 transcription.A Representative western blots of HeLa cells after transfection with HPV18 E6/E7 targeting siRNA. Cell lysates were probed for TAZ, YAP, HPV18 E6 and HPV18 E7 expression. GAPDH was used as a loading control (n = 4). B RT-qPCR analysis of WWTR1 expression in HeLa cells after transfection with HPV18 E6/E7 targeting siRNA (n = 3). U6 was used as a loading control. CWWTR1-promoter luciferase assay in HeLa cells after transfection with HPV18 E6/E7 targeting siRNA (n = 3). D Representative western blot of C33A cell lysates stably expressing either HA-HPV18 E6 or HA-HPV18 E7 probed for TAZ, YAP, HPV18 E6, HPV18 E7 and the loading control GAPDH (n = 3). E RT-qPCR analysis of WWTR1 expression in C33A cells stably expressing either HA-HPV18 E6 or HA-HPV18 E7 (n = 3). U6 was used as a loading control. FWWTR1-promoter luciferase assay in C33A cells stably expressing either HA-HPV18 E6 or HA-HPV18 E7 (n = 3). G Representative western blot of C33A cell lysate stably expressing either HA-HPV18 E6 or HA-HPV18 E7 probed for TAZ, YAP, HPV16 E7, HPV18 E7 and GAPDH (n = 3). H RT-qPCR analysis of WWTR1 expression in C33A cells stably expressing either HA-HPV16 E7 or HA-HPV18 E7 (n = 4). U6 was used as a loading control. IWWTR1-promoter luciferase assay in C33A cells stably expressing either HA-HPV16 E7 or HA-HPV18 E7 (n = 3). Error bars represent the mean +/- standard deviation of a minimum of three biological repeats. *P < 0.05, **P < 0.01, ***P < 0.005 (two-tailed, unpaired Student’s t-test). Source data are provided as a Source Data file.
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Collected and cropped from Nature Communications by CiteAb, provided under a CC-BY license
Image 1 of 32
In Nat Commun on 10 July 2024 by Patterson, M. R., Cogan, J. A., et al.
Fig.5.G
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YAP and TAZ are oncogenic in HPV18 + , but not HPV16+ cervical cancer cell lines.A RT-qPCR analysis of WWTR1 and YAP1 expression in TAZ, YAP or YAP/TAZ (Y/T) KD HeLa cells (n = 3). U6 was used as a loading control. B Representative western blots of TAZ, YAP and loading control GAPDH from TAZ, YAP...
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YAP and TAZ are oncogenic in HPV18 + , but not HPV16+ cervical cancer cell lines.A RT-qPCR analysis of WWTR1 and YAP1 expression in TAZ, YAP or YAP/TAZ (Y/T) KD HeLa cells (n = 3). U6 was used as a loading control. B Representative western blots of TAZ, YAP and loading control GAPDH from TAZ, YAP or YAP/TAZ (Y/T) KD HeLa cell lysates (n = 3). C Growth curve analysis of TAZ, YAP or YAP/TAZ (Y/T) KD HeLa cells (n = 3). D Colony formation assay of TAZ, YAP or YAP/TAZ (Y/T) KD HeLa cells (n = 3). E Soft agar assay of TAZ, YAP or YAP/TAZ (Y/T) KD HeLa cells (n = 3). F RT-qPCR analysis of WWTR1 or YAP1 expression in TAZ, YAP or YAP/TAZ (Y/T) KD SiHa cells (n = 3). U6 was used as a loading control. G Representative western blot of YAP, TAZ and GAPDH from TAZ, YAP or YAP/ TAZ (Y/T) KD SiHa cell lysates (n = 3). H Growth curve analysis of TAZ, YAP or YAP/TAZ (Y/T) KD SiHa cells (n = 3). I Colony formation assay of TAZ, YAP or YAP/TAZ (Y/T) KD SiHa cells (n = 3). J Soft agar assay of TAZ, YAP or YAP/TAZ (Y/T) KD SiHa cells (n = 3). Error bars represent the mean +/- standard deviation of a minimum of three biological repeats. *P < 0.05, **P < 0.01, ***P < 0.005 (two-tailed, unpaired Student’s t-test). Source data are provided as a Source Data file.
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Collected and cropped from Nature Communications by CiteAb, provided under a CC-BY license
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In Nat Commun on 10 July 2024 by Patterson, M. R., Cogan, J. A., et al.
Fig.9.A
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Homo sapiens (Human)
TOGARAM2 contributes towards TAZ-driven oncogenicity.A Representative western blots of TAZ KD HeLa cell lysates transfected with pcDNA3 or FLAG-TOGARAM2 probed for TAZ, FLAG and GAPDH loading control (n = 3). B Growth curve analysis of TAZ KD HeLa cell lysates transfected with pcDNA3 or FLAG-TOGA...
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TOGARAM2 contributes towards TAZ-driven oncogenicity.A Representative western blots of TAZ KD HeLa cell lysates transfected with pcDNA3 or FLAG-TOGARAM2 probed for TAZ, FLAG and GAPDH loading control (n = 3). B Growth curve analysis of TAZ KD HeLa cell lysates transfected with pcDNA3 or FLAG-TOGARAM2 (n = 3). C Colony formation assay of TAZ KD HeLa cell lysates transfected with pcDNA3 or FLAG-TOGARAM2 (n = 4). D Soft agar assay of TAZ KD HeLa cell lysates transfected with pcDNA3 or FLAG-TOGARAM2 (n = 4). E Transwell migration assay of TAZ KD HeLa cells transfected with pcDNA3 or FLAG-TOGARAM2 (n = 3). F Cell Invasion assay of TAZ KD HeLa cells transfected with pcDNA3 or FLAG-TOGARAM2 (n = 4). G Immunofluorescence microscopy analysis of Rhodamine-Phalloidin stained (red) TAZ KD HeLa cells transfected with pcDNA3 or FLAG-TOGARAM2. DAPI stained nuclei (blue). Scale bar 10 µm. H Measurement of filopodia length in TAZ KD HeLa cells transfected with pcDNA3 or FLAG-TOGARAM2 (n = 10). I Average number of filopodia per cell in TAZ KD HeLa cells transfected with pcDNA3 or FLAG-TOGARAM2 (n = 10). Error bars represent the mean +/- standard deviation of a minimum of three biological repeats. *P < 0.05, **P < 0.01, ***P < 0.005 (two-tailed, unpaired Student’s t-test). Source data are provided as a Source Data file.
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Collected and cropped from Nature Communications by CiteAb, provided under a CC-BY license
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In Front Oncol on 22 May 2024 by Choi, H. S., Jang, H. J., et al.
Fig.6.A
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Homo sapiens (Human)
Semiquantitative immunoblotting of RhoA-siRNA-treated MDA-MB-231 cells and cell migration assay in MDA-MB-231 cells treated with control-siRNA, TAZ-siRNA, or RhoA-siRNA, respectively. (A–D) Changes in protein abundance of RhoA and TAZ in MDA-MB-231 cell treated with control-siRNA or RhoA-siRNA fo...
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Semiquantitative immunoblotting of RhoA-siRNA-treated MDA-MB-231 cells and cell migration assay in MDA-MB-231 cells treated with control-siRNA, TAZ-siRNA, or RhoA-siRNA, respectively. (A–D) Changes in protein abundance of RhoA and TAZ in MDA-MB-231 cell treated with control-siRNA or RhoA-siRNA for 24 hours. *P < 0.05, when compared with control-siRNA. The immunoblots were reacted with antibodies against RhoA (~22kDa), TAZ (~49 kDa), and β-actin (~42 kDa). n indicates the number of cell preparation in each group. (D, E) Transwell cell migration assay of MDA-MB-231 cells treated with control-siRNA, TAZ-siRNA, or RhoA-siRNA treatment. The numbers of migrated cells were counted in the randomly selected fields (x100) per well. n indicates the number of randomly selected fields in each group. *P < 0.05.
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Collected and cropped from Frontiers in Oncology by CiteAb, provided under a CC-BY license
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In Front Oncol on 22 May 2024 by Choi, H. S., Jang, H. J., et al.
Fig.3.B
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Semiquantitative immunoblotting for the studies of TGF-β treatment. (A, C–H) Semiquantitative immunoblotting of TAZ, E-cadherin, occludin, fibronectin, N-cadherin, vimentin, and β-actin in MDA-MB-231 cells treated with vehicle or TGF-β (5 ng/ml) under control-siRNA treatment. The immunoblots were...
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Semiquantitative immunoblotting for the studies of TGF-β treatment. (A, C–H) Semiquantitative immunoblotting of TAZ, E-cadherin, occludin, fibronectin, N-cadherin, vimentin, and β-actin in MDA-MB-231 cells treated with vehicle or TGF-β (5 ng/ml) under control-siRNA treatment. The immunoblots were reacted with antibodies against TAZ (~49 kDa), E-cadherin (~135 kDa), occludin (~65 kDa), fibronectin (~262 kDa), N-cadherin (~125 kDa), vimentin (~57 kDa), and β-actin (~42 kDa). (B, I–N) Semiquantitative immunoblotting of TAZ, E-cadherin, occludin, fibronectin, N-cadherin, vimentin, and β-actin in MDA-MB-231 cells treated with vehicle or TGF-β (5 ng/ml) under TAZ-siRNA treatment. *P < 0.05, when compared with vehicle. n indicates the number of cell preparation in each group. Transwell cell migration assay of MDA-MB-231 cells treated with vehicle or TGF-β (5 ng/ml) under control-siRNA (O) or TAZ-siRNA treatment (P). (Q) The numbers of migrated cells were counted in the randomly selected fields (x100) per well. n indicates the number of randomly selected fields in each group. *P < 0.05.
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Collected and cropped from Frontiers in Oncology by CiteAb, provided under a CC-BY license
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In Front Oncol on 22 May 2024 by Choi, H. S., Jang, H. J., et al.
Fig.1.B
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Semiquantitative immunoblotting of TAZ and cell migration assay in breast cancer cells (MCF-7 and MDA-MB-231) treated with control-siRNA or TAZ-siRNA. (A–D) Changes in protein abundance of TAZ in breast cancer cells treated with control-siRNA or TAZ-siRNA for 24 hours. *P < 0.05, when compared wi...
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Semiquantitative immunoblotting of TAZ and cell migration assay in breast cancer cells (MCF-7 and MDA-MB-231) treated with control-siRNA or TAZ-siRNA. (A–D) Changes in protein abundance of TAZ in breast cancer cells treated with control-siRNA or TAZ-siRNA for 24 hours. *P < 0.05, when compared with control-siRNA. The immunoblots were reacted with antibodies against TAZ (~49 kDa) and β-actin (~42 kDa). n indicates the number of cell preparation in each group. (E–H) Transwell cell migration assay of breast cancer cells treated with control-siRNA or TAZ-siRNA. The numbers of migrated cells were counted in the randomly selected fields (x100) per well. n indicates the number of randomly selected fields in each group. *P < 0.05, when compared with control-siRNA. (I–L) Scratch wound healing assay. *P < 0.05, when compared with control-siRNA. n indicates the number of wells containing cell treated with control-siRNA or TAZ-siRNA.
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Collected and cropped from Frontiers in Oncology by CiteAb, provided under a CC-BY license
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In Front Oncol on 22 May 2024 by Choi, H. S., Jang, H. J., et al.
Fig.1.A
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Homo sapiens (Human)
Semiquantitative immunoblotting of TAZ and cell migration assay in breast cancer cells (MCF-7 and MDA-MB-231) treated with control-siRNA or TAZ-siRNA. (A–D) Changes in protein abundance of TAZ in breast cancer cells treated with control-siRNA or TAZ-siRNA for 24 hours. *P < 0.05, when compared wi...
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Semiquantitative immunoblotting of TAZ and cell migration assay in breast cancer cells (MCF-7 and MDA-MB-231) treated with control-siRNA or TAZ-siRNA. (A–D) Changes in protein abundance of TAZ in breast cancer cells treated with control-siRNA or TAZ-siRNA for 24 hours. *P < 0.05, when compared with control-siRNA. The immunoblots were reacted with antibodies against TAZ (~49 kDa) and β-actin (~42 kDa). n indicates the number of cell preparation in each group. (E–H) Transwell cell migration assay of breast cancer cells treated with control-siRNA or TAZ-siRNA. The numbers of migrated cells were counted in the randomly selected fields (x100) per well. n indicates the number of randomly selected fields in each group. *P < 0.05, when compared with control-siRNA. (I–L) Scratch wound healing assay. *P < 0.05, when compared with control-siRNA. n indicates the number of wells containing cell treated with control-siRNA or TAZ-siRNA.
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Collected and cropped from Frontiers in Oncology by CiteAb, provided under a CC-BY license
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In Front Oncol on 22 May 2024 by Choi, H. S., Jang, H. J., et al.
Fig.4.A
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Semiquantitative immunoblotting for Rho family GTPases. Changes in Rho GTPase protein abundance in breast cancer cells treated with control-siRNA or TAZ-siRNA. (A–H) Semiquantitative immunoblotting of TAZ, YAP, RhoA, RhoC, ROCK, Cdc42, Rac1, and β-actin in MDA-MB-231 cells treated with control-si...
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Semiquantitative immunoblotting for Rho family GTPases. Changes in Rho GTPase protein abundance in breast cancer cells treated with control-siRNA or TAZ-siRNA. (A–H) Semiquantitative immunoblotting of TAZ, YAP, RhoA, RhoC, ROCK, Cdc42, Rac1, and β-actin in MDA-MB-231 cells treated with control-siRNA or TAZ-siRNA. The immunoblots were reacted with antibodies against TAZ (~49 kDa), RhoA (~22 kDa), RhoC (~21 kDa), ROCK (~158 kDa), Cdc42 (~21 kDa), Rac1 (~21 kDa), and β-actin (~42 kDa).
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Collected and cropped from Frontiers in Oncology by CiteAb, provided under a CC-BY license
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In Front Oncol on 22 May 2024 by Choi, H. S., Jang, H. J., et al.
Fig.3.A
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WB
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Homo sapiens (Human)
Semiquantitative immunoblotting for the studies of TGF-β treatment. (A, C–H) Semiquantitative immunoblotting of TAZ, E-cadherin, occludin, fibronectin, N-cadherin, vimentin, and β-actin in MDA-MB-231 cells treated with vehicle or TGF-β (5 ng/ml) under control-siRNA treatment. The immunoblots were...
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Semiquantitative immunoblotting for the studies of TGF-β treatment. (A, C–H) Semiquantitative immunoblotting of TAZ, E-cadherin, occludin, fibronectin, N-cadherin, vimentin, and β-actin in MDA-MB-231 cells treated with vehicle or TGF-β (5 ng/ml) under control-siRNA treatment. The immunoblots were reacted with antibodies against TAZ (~49 kDa), E-cadherin (~135 kDa), occludin (~65 kDa), fibronectin (~262 kDa), N-cadherin (~125 kDa), vimentin (~57 kDa), and β-actin (~42 kDa). (B, I–N) Semiquantitative immunoblotting of TAZ, E-cadherin, occludin, fibronectin, N-cadherin, vimentin, and β-actin in MDA-MB-231 cells treated with vehicle or TGF-β (5 ng/ml) under TAZ-siRNA treatment. *P < 0.05, when compared with vehicle. n indicates the number of cell preparation in each group. Transwell cell migration assay of MDA-MB-231 cells treated with vehicle or TGF-β (5 ng/ml) under control-siRNA (O) or TAZ-siRNA treatment (P). (Q) The numbers of migrated cells were counted in the randomly selected fields (x100) per well. n indicates the number of randomly selected fields in each group. *P < 0.05.
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Collected and cropped from Frontiers in Oncology by CiteAb, provided under a CC-BY license
Image 1 of 32
In Front Oncol on 22 May 2024 by Choi, H. S., Jang, H. J., et al.
Fig.2.A
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Homo sapiens (Human)
Semiquantitative immunoblotting for the studies of EMT. (A–G) Semiquantitative immunoblotting of TAZ, E-cadherin, occludin, fibronectin, N-cadherin, vimentin and β-actin in MDA-MB-231 cells treated with control-siRNA or TAZ-siRNA. The immunoblots were reacted with antibodies against TAZ (~49 kDa)...
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Semiquantitative immunoblotting for the studies of EMT. (A–G) Semiquantitative immunoblotting of TAZ, E-cadherin, occludin, fibronectin, N-cadherin, vimentin and β-actin in MDA-MB-231 cells treated with control-siRNA or TAZ-siRNA. The immunoblots were reacted with antibodies against TAZ (~49 kDa), E-cadherin (~135 kDa), occludin (~65 kDa), fibronectin (~262 kDa), N-cadherin (~125 kDa), vimentin (~57 kDa), and β-actin (~42 kDa). *P < 0.05, when compared with control-siRNA. n indicates the number of cell preparation in each group.
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Collected and cropped from Frontiers in Oncology by CiteAb, provided under a CC-BY license
Image 1 of 32
In Cell Death Dis on 9 July 2020 by Wang, Y., Liao, R., et al.
Fig.4.D
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PAR1 alters the expression of EMT markers and inhibits the Hippo pathway.a Expression of PAR1 was analyzed by quantitative real-time PCR in MDA-MB-231 and Hs578T cells with stable control vector or knockdown of PAR1 expression. Data are presented as mean ± SD of three separate experiments. b Expr...
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PAR1 alters the expression of EMT markers and inhibits the Hippo pathway.a Expression of PAR1 was analyzed by quantitative real-time PCR in MDA-MB-231 and Hs578T cells with stable control vector or knockdown of PAR1 expression. Data are presented as mean ± SD of three separate experiments. b Expression of PAR1, E-cadherin, and Vimentin was assessed by western blotting in MDA-MB-231 and Hs578T cells with stable control vector or knockdown of PAR1 expression, and actin was served as a loading control. Representative images were presented from three independent experiments. c The morphologic changes of MDA-MB-231 and Hs578T cells with stable control vector or knockdown of PAR1 expression were shown in representative images. Scale bars, 100 μm. d Expression of YAP, TAZ, and CTGF was analyzed by western blotting in MDA-MB-231 and Hs578T cells with stable control vector or knockdown of PAR1 expression. Representative images were presented from three independent experiments. e Expression of PAR1, YAP, and TAZ was assessed by western blot in T47D cells transfected with control vector or PAR1-expressing vector, and actin was served as a loading control. Representative images were presented from three independent experiments. f The CTGF reporter activity was examined in MDA-MB-231 cells with control vector or knockdown of PAR1 expression. After 48 h, luciferase activities were determined and normalized (mean ± SD of three separate experiments) *P < 0.01 by Student’s t test. g T47D-Twist and MDA-MB-231 cells were serum-starved for 24 h, and then treated with indicated concentration of vorapaxar. Expression of PAR1, YAP, and TAZ was assessed by western blotting, and actin was served as a loading control. Representative images were presented from three independent experiments. h Expression of Foxc2 was analyzed by quantitative real-time PCR in MDA-MB-231 and Hs578T cells with stable control vector or knockdown of PAR1 expression. Data are presented as mean ± SD of three separate experiments.
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Collected and cropped from Cell Death & Disease by CiteAb, provided under a CC-BY license
Image 1 of 32
In Cell Death Dis on 9 July 2020 by Wang, Y., Liao, R., et al.
Fig.4.E
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PAR1 alters the expression of EMT markers and inhibits the Hippo pathway.a Expression of PAR1 was analyzed by quantitative real-time PCR in MDA-MB-231 and Hs578T cells with stable control vector or knockdown of PAR1 expression. Data are presented as mean ± SD of three separate experiments. b Expr...
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PAR1 alters the expression of EMT markers and inhibits the Hippo pathway.a Expression of PAR1 was analyzed by quantitative real-time PCR in MDA-MB-231 and Hs578T cells with stable control vector or knockdown of PAR1 expression. Data are presented as mean ± SD of three separate experiments. b Expression of PAR1, E-cadherin, and Vimentin was assessed by western blotting in MDA-MB-231 and Hs578T cells with stable control vector or knockdown of PAR1 expression, and actin was served as a loading control. Representative images were presented from three independent experiments. c The morphologic changes of MDA-MB-231 and Hs578T cells with stable control vector or knockdown of PAR1 expression were shown in representative images. Scale bars, 100 μm. d Expression of YAP, TAZ, and CTGF was analyzed by western blotting in MDA-MB-231 and Hs578T cells with stable control vector or knockdown of PAR1 expression. Representative images were presented from three independent experiments. e Expression of PAR1, YAP, and TAZ was assessed by western blot in T47D cells transfected with control vector or PAR1-expressing vector, and actin was served as a loading control. Representative images were presented from three independent experiments. f The CTGF reporter activity was examined in MDA-MB-231 cells with control vector or knockdown of PAR1 expression. After 48 h, luciferase activities were determined and normalized (mean ± SD of three separate experiments) *P < 0.01 by Student’s t test. g T47D-Twist and MDA-MB-231 cells were serum-starved for 24 h, and then treated with indicated concentration of vorapaxar. Expression of PAR1, YAP, and TAZ was assessed by western blotting, and actin was served as a loading control. Representative images were presented from three independent experiments. h Expression of Foxc2 was analyzed by quantitative real-time PCR in MDA-MB-231 and Hs578T cells with stable control vector or knockdown of PAR1 expression. Data are presented as mean ± SD of three separate experiments.
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Collected and cropped from Cell Death & Disease by CiteAb, provided under a CC-BY license
Image 1 of 32
In Nat Commun on 23 November 2018 by Kofler, M., Speight, P., et al.
Fig.1.B
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Sus scrofa (Wild pig)
Large tags suppress nuclear accumulation of TAZ under steady-state conditions. a TAZ and a control (mCherry, red circle) were fused to increasing numbers of mCitrine molecules (yellow circles). Molecular weights are indicated. b Western blot analysis of transiently over-expressed TAZ constructs i...
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Large tags suppress nuclear accumulation of TAZ under steady-state conditions. a TAZ and a control (mCherry, red circle) were fused to increasing numbers of mCitrine molecules (yellow circles). Molecular weights are indicated. b Western blot analysis of transiently over-expressed TAZ constructs in LLC-PK1 cells, using a TAZ-specific antibody. Endogenous TAZ is marked by an arrowhead. An uncropped version is shown in Supplementary Figure 1A. c Fluorescence microscopy images of LLC-PK1 cells transiently expressing control or TAZ 4SA constructs with varying tag-sizes (numbers of mCitrine units). 5C-control-SV40 additionally comprises residues 114–135 of simian virus 40 (SV40) large tumor antigen that mediate strong nuclear import. This construct demonstrates that the 5C-tag does not abolish mediated import and serves as positive control for nuclear localization. Images depict mCitrine fluorescence. Scale bar represents 50 μm. Representative images of one experiment. Number of repeats: n = 3. d Visual quantification of cells with nuclear enrichment of control and TAZ 4SA constructs. Cells were scored as “nuclear” (nuc.) if nuclear fluorescence was visibly higher than cytoplasmic fluorescence. For each measurement, at least 100 cells were analysed. See also Supplementary Figure 3B. Data are represented as bar diagram, n = 7. Throughout this work, data from visual analyses are depicted in the same way. e Automated analysis of the localization of indicated constructs in cells, shown in (c), using the ImageXpress platform. For each construct, nucleocytoplasmic fluorescence ratios (RN/C) from more than 100 cells were obtained from the inbuilt automated cell-recognition algorithm and the medians of these ratio distributions were calculated. Depicted histograms are averages, n = 3. f Comparison of median RN/C, determined in (e). Dotted and dashed lines represent RN/C thresholds for the classification of nuclear exclusion and accumulation, respectively, as described in Supplementary Figure 3C. In the following, all automated analyses are presented as scatter charts. Data are represented as means ± SD. *p < 0.05, **p < 0.01; Student’s t-test
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Collected and cropped from Nature Communications by CiteAb, provided under a CC-BY license
Image 1 of 32
In Nat Commun on 23 November 2018 by Kofler, M., Speight, P., et al.
Fig.8.H
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Sus scrofa (Wild pig)
The TBD comprises a NES within the TEAD-binding site. a Sequence alignment of TBDs from TAZ homologs, as in Fig. 5b. Positively charged residues are depicted in red. Filled and open boxes (above alignment) highlight important or dispensable positions for TEAD binding, respectively25. TEAD-binding...
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The TBD comprises a NES within the TEAD-binding site. a Sequence alignment of TBDs from TAZ homologs, as in Fig. 5b. Positively charged residues are depicted in red. Filled and open boxes (above alignment) highlight important or dispensable positions for TEAD binding, respectively25. TEAD-binding sites are boxed in yellow54, 55. Mutations used in this study are shown below. b Nuclear enrichment of 1C-TBD mutants, presented as mean ± SD, n = 3. c Impact of TBD mutations on TEAD binding. 5C-TAZ constructs and the Myc-TAZ binding domain of TEAD1 (TAZBD) were co-expressed. 5C-constructs from lysates were immunoprecipitated with GFP-trap beads. TAZ and Myc-TAZBD were detected in lysate (input) and immunoprecipitates (5C-IP) by western analysis using TAZ- and Myc-specific antibodies, respectively. d RIS’C model showing rapamycin-induced cytoplasmic entrapment of 1C-FRB-TAZ 4SA F52A (1C-FRB-TAZ*) by myristoylated-2xFKBP-mCherry. e RIS’C assay using live-cell imaging. Cells expressing Myr-2xFKBP-mCherry and 1C-FRB-TAZ* or 1C-FRB-TAZ*-M2 were imaged confocally. After the first image (t = 0 min), rapamycin was added and further images were taken. Scale bar: 10 μm. Average RN/C values ± SEM are plotted against time. 1C-FRB-TAZ*, n = 43 cells, 1C-FRB-TAZ*-M2, n = 29 cells. Significant differences between the two probes are indicated in time-intervals. Fits represent an exponential decay to minimum function. f LMB sensitivity of nuclear export. Experiments and fits were performed as in (e), with or without additional LMB treatment (open and closed circles) for 80 min. Median RN/C values of 1C-FRB-TAZ* (blue) and 1C-FRB-TAZ*-M2 (red) are presented as means ± SD, n = 3. Curves correspond to an exponential decay function. In the presence of LMB, the decays of nuclear accumulation of the constructs are indistinguishable (single fit; black dotted curve). Blue and red asterisks indicate minimum significance levels of 1C-FRB-TAZ* relative to all other conditions and 1C-FRB-TAZ*-M2 relative to LMB-treated samples, respectively. g Nuclear enrichment of 1C-TAZ constructs with mutations in the TBD, presented as means ± SD, n = 4. h Competition between TAZBD and 14–3–3 for TAZ binding. 5C-control and 5C-TAZ constructs were co-expressed with or without Myc-TAZBD, immunoprecipitated with GFP-trap beads, and analyzed by western blotting using TAZ-, Myc-, and 14–3–3 specific antibodies. Note weak, unspecific binding of Myc-TAZBD to 5C-control. Uncropped versions of blots are shown in Supplementary Figure 1C and 1D. *p < 0.05, **p < 0.01, ***p < 0.001; Student’s t-test
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Collected and cropped from Nature Communications by CiteAb, provided under a CC-BY license
Image 1 of 32
In Nat Commun on 23 November 2018 by Kofler, M., Speight, P., et al.
Fig.8.C
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Sus scrofa (Wild pig)
The TBD comprises a NES within the TEAD-binding site. a Sequence alignment of TBDs from TAZ homologs, as in Fig. 5b. Positively charged residues are depicted in red. Filled and open boxes (above alignment) highlight important or dispensable positions for TEAD binding, respectively25. TEAD-binding...
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The TBD comprises a NES within the TEAD-binding site. a Sequence alignment of TBDs from TAZ homologs, as in Fig. 5b. Positively charged residues are depicted in red. Filled and open boxes (above alignment) highlight important or dispensable positions for TEAD binding, respectively25. TEAD-binding sites are boxed in yellow54, 55. Mutations used in this study are shown below. b Nuclear enrichment of 1C-TBD mutants, presented as mean ± SD, n = 3. c Impact of TBD mutations on TEAD binding. 5C-TAZ constructs and the Myc-TAZ binding domain of TEAD1 (TAZBD) were co-expressed. 5C-constructs from lysates were immunoprecipitated with GFP-trap beads. TAZ and Myc-TAZBD were detected in lysate (input) and immunoprecipitates (5C-IP) by western analysis using TAZ- and Myc-specific antibodies, respectively. d RIS’C model showing rapamycin-induced cytoplasmic entrapment of 1C-FRB-TAZ 4SA F52A (1C-FRB-TAZ*) by myristoylated-2xFKBP-mCherry. e RIS’C assay using live-cell imaging. Cells expressing Myr-2xFKBP-mCherry and 1C-FRB-TAZ* or 1C-FRB-TAZ*-M2 were imaged confocally. After the first image (t = 0 min), rapamycin was added and further images were taken. Scale bar: 10 μm. Average RN/C values ± SEM are plotted against time. 1C-FRB-TAZ*, n = 43 cells, 1C-FRB-TAZ*-M2, n = 29 cells. Significant differences between the two probes are indicated in time-intervals. Fits represent an exponential decay to minimum function. f LMB sensitivity of nuclear export. Experiments and fits were performed as in (e), with or without additional LMB treatment (open and closed circles) for 80 min. Median RN/C values of 1C-FRB-TAZ* (blue) and 1C-FRB-TAZ*-M2 (red) are presented as means ± SD, n = 3. Curves correspond to an exponential decay function. In the presence of LMB, the decays of nuclear accumulation of the constructs are indistinguishable (single fit; black dotted curve). Blue and red asterisks indicate minimum significance levels of 1C-FRB-TAZ* relative to all other conditions and 1C-FRB-TAZ*-M2 relative to LMB-treated samples, respectively. g Nuclear enrichment of 1C-TAZ constructs with mutations in the TBD, presented as means ± SD, n = 4. h Competition between TAZBD and 14–3–3 for TAZ binding. 5C-control and 5C-TAZ constructs were co-expressed with or without Myc-TAZBD, immunoprecipitated with GFP-trap beads, and analyzed by western blotting using TAZ-, Myc-, and 14–3–3 specific antibodies. Note weak, unspecific binding of Myc-TAZBD to 5C-control. Uncropped versions of blots are shown in Supplementary Figure 1C and 1D. *p < 0.05, **p < 0.01, ***p < 0.001; Student’s t-test
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Collected and cropped from Nature Communications by CiteAb, provided under a CC-BY license
Image 1 of 32
In Nat Commun on 29 August 2018 by Gill, M. K., Christova, T., et al.
Fig.4.G
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NUAK2 activity is involved in serum-induced or LPA-induced YAP/TAZ dephosphorylation and activation. a, b FBS and LPA activate YAP/TAZ and induce NUAK2 expression. YAP/TAZ phosphorylation status was monitored using Phos-Tag gels (a), while NUAK2 and ANKRD1 mRNA expression was determined by qPCR a...
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NUAK2 activity is involved in serum-induced or LPA-induced YAP/TAZ dephosphorylation and activation. a, b FBS and LPA activate YAP/TAZ and induce NUAK2 expression. YAP/TAZ phosphorylation status was monitored using Phos-Tag gels (a), while NUAK2 and ANKRD1 mRNA expression was determined by qPCR at the indicated times. Expression data are plotted as the mean ± SD (n = 3). c–h WZ4003 (10 μM) blocks serum/LPA-induced NUAK2 and ANKRD1 mRNA expression. FBS-induced or LPA-induced YAP/TAZ dephosphorylation (e and g), or NUAK2 and ANKRD1 mRNA expression (c, f, h) was monitored at the indicated times (d). Expression data are plotted as the mean ± range for a representative experiment (c, f, h)
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Collected and cropped from Nature Communications by CiteAb, provided under a CC-BY license
Image 1 of 32
In Nat Commun on 29 August 2018 by Gill, M. K., Christova, T., et al.
Fig.4.A
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NUAK2 activity is involved in serum-induced or LPA-induced YAP/TAZ dephosphorylation and activation. a, b FBS and LPA activate YAP/TAZ and induce NUAK2 expression. YAP/TAZ phosphorylation status was monitored using Phos-Tag gels (a), while NUAK2 and ANKRD1 mRNA expression was determined by qPCR a...
more
NUAK2 activity is involved in serum-induced or LPA-induced YAP/TAZ dephosphorylation and activation. a, b FBS and LPA activate YAP/TAZ and induce NUAK2 expression. YAP/TAZ phosphorylation status was monitored using Phos-Tag gels (a), while NUAK2 and ANKRD1 mRNA expression was determined by qPCR at the indicated times. Expression data are plotted as the mean ± SD (n = 3). c–h WZ4003 (10 μM) blocks serum/LPA-induced NUAK2 and ANKRD1 mRNA expression. FBS-induced or LPA-induced YAP/TAZ dephosphorylation (e and g), or NUAK2 and ANKRD1 mRNA expression (c, f, h) was monitored at the indicated times (d). Expression data are plotted as the mean ± range for a representative experiment (c, f, h)
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Collected and cropped from Nature Communications by CiteAb, provided under a CC-BY license
Image 1 of 32
In Nat Commun on 29 August 2018 by Gill, M. K., Christova, T., et al.
Fig.4.E
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NUAK2 activity is involved in serum-induced or LPA-induced YAP/TAZ dephosphorylation and activation. a, b FBS and LPA activate YAP/TAZ and induce NUAK2 expression. YAP/TAZ phosphorylation status was monitored using Phos-Tag gels (a), while NUAK2 and ANKRD1 mRNA expression was determined by qPCR a...
more
NUAK2 activity is involved in serum-induced or LPA-induced YAP/TAZ dephosphorylation and activation. a, b FBS and LPA activate YAP/TAZ and induce NUAK2 expression. YAP/TAZ phosphorylation status was monitored using Phos-Tag gels (a), while NUAK2 and ANKRD1 mRNA expression was determined by qPCR at the indicated times. Expression data are plotted as the mean ± SD (n = 3). c–h WZ4003 (10 μM) blocks serum/LPA-induced NUAK2 and ANKRD1 mRNA expression. FBS-induced or LPA-induced YAP/TAZ dephosphorylation (e and g), or NUAK2 and ANKRD1 mRNA expression (c, f, h) was monitored at the indicated times (d). Expression data are plotted as the mean ± range for a representative experiment (c, f, h)
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Collected and cropped from Nature Communications by CiteAb, provided under a CC-BY license
Image 1 of 32
In Nat Commun on 13 March 2018 by Azad, T., Janse van Rensburg, H. J., et al.
Fig.5.A
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VEGFR regulates angiogenesis and tumor VM through YAP/TAZ in vitro. a Transient knockdown of YAP and/or TAZ in MCF10A overexpressing VEGFR2 decreases expression of ANG-2 and CYR61. Western blotting exposures indicate relative expression of YAP and TAZ. b, c VEGFR2 overexpression and VEGF treatmen...
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VEGFR regulates angiogenesis and tumor VM through YAP/TAZ in vitro. a Transient knockdown of YAP and/or TAZ in MCF10A overexpressing VEGFR2 decreases expression of ANG-2 and CYR61. Western blotting exposures indicate relative expression of YAP and TAZ. b, c VEGFR2 overexpression and VEGF treatment increases tube formation by MCF10A through YAP/TAZ. YAP and/or TAZ were transiently knocked down by siRNA in MCF10A stably overexpressing VEGFR2 and subjected to tube-formation assay on Matrigel 48 h after transfection alongside wild type MCF10A. For some conditions, cells were stimulated with 100 ng ml−1 VEGF or were treated with 100 nM verteporfin for the duration of the tube formation assay. Representative images are shown in b. Scale bar denotes 200 μm. Total tube formation was quantified in c (n = 3). *p < 0.05 in two-sample unpaired t-test. d–l YAP/TAZ are critical for VEGF-induced angiogenesis in HUVEC (d–f) and BOEC (g–i), as well as for vasculogenic mimicry in MDA-MB231 (j–l). d, g, j YAP and/or TAZ were transiently knocked down by siRNA in each cell line which reduced ANG-2 and CYR61 expression. Western blotting exposures indicate relative expression of YAP and TAZ for each cell line. e, h, k Representative images of tube formation on Matrigel assessed 48 h after transfection. For some conditions, cells were stimulated with 100 ng ml−1 VEGF or were treated with 100 nM verteporfin for the duration of the tube formation assay. Scale bar denotes 200 μm. Total tube formation was quantified in f, i, l (n = 3). *p < 0.05 in two-sample unpaired t-test. m, n Exogenous CYR61 and ANG-2 can partially rescue tube formation in YAP/TAZ knockdown HUVEC. YAP and/or TAZ were transiently knocked down by siRNA in HUVEC. Tube formation on Matrigel was assessed 48 h after transfection. For some conditions, cells were stimulated with 100 ng ml−1 VEGF, 200 ng ml−1 CYR61, and/or 200 ng ml−1ANG-2 for the duration of the tube formation assay. Scale bar denotes 200 μm. Representative images are shown in m and total tube formation was quantified in n. (n = 3). All data are represented as mean ± SD
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Collected and cropped from Nature Communications by CiteAb, provided under a CC-BY license
Image 1 of 32
In Nat Commun on 13 March 2018 by Azad, T., Janse van Rensburg, H. J., et al.
Fig.5.D
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VEGFR regulates angiogenesis and tumor VM through YAP/TAZ in vitro. a Transient knockdown of YAP and/or TAZ in MCF10A overexpressing VEGFR2 decreases expression of ANG-2 and CYR61. Western blotting exposures indicate relative expression of YAP and TAZ. b, c VEGFR2 overexpression and VEGF treatmen...
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VEGFR regulates angiogenesis and tumor VM through YAP/TAZ in vitro. a Transient knockdown of YAP and/or TAZ in MCF10A overexpressing VEGFR2 decreases expression of ANG-2 and CYR61. Western blotting exposures indicate relative expression of YAP and TAZ. b, c VEGFR2 overexpression and VEGF treatment increases tube formation by MCF10A through YAP/TAZ. YAP and/or TAZ were transiently knocked down by siRNA in MCF10A stably overexpressing VEGFR2 and subjected to tube-formation assay on Matrigel 48 h after transfection alongside wild type MCF10A. For some conditions, cells were stimulated with 100 ng ml−1 VEGF or were treated with 100 nM verteporfin for the duration of the tube formation assay. Representative images are shown in b. Scale bar denotes 200 μm. Total tube formation was quantified in c (n = 3). *p < 0.05 in two-sample unpaired t-test. d–l YAP/TAZ are critical for VEGF-induced angiogenesis in HUVEC (d–f) and BOEC (g–i), as well as for vasculogenic mimicry in MDA-MB231 (j–l). d, g, j YAP and/or TAZ were transiently knocked down by siRNA in each cell line which reduced ANG-2 and CYR61 expression. Western blotting exposures indicate relative expression of YAP and TAZ for each cell line. e, h, k Representative images of tube formation on Matrigel assessed 48 h after transfection. For some conditions, cells were stimulated with 100 ng ml−1 VEGF or were treated with 100 nM verteporfin for the duration of the tube formation assay. Scale bar denotes 200 μm. Total tube formation was quantified in f, i, l (n = 3). *p < 0.05 in two-sample unpaired t-test. m, n Exogenous CYR61 and ANG-2 can partially rescue tube formation in YAP/TAZ knockdown HUVEC. YAP and/or TAZ were transiently knocked down by siRNA in HUVEC. Tube formation on Matrigel was assessed 48 h after transfection. For some conditions, cells were stimulated with 100 ng ml−1 VEGF, 200 ng ml−1 CYR61, and/or 200 ng ml−1ANG-2 for the duration of the tube formation assay. Scale bar denotes 200 μm. Representative images are shown in m and total tube formation was quantified in n. (n = 3). All data are represented as mean ± SD
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Collected and cropped from Nature Communications by CiteAb, provided under a CC-BY license
Image 1 of 32
In Nat Commun on 13 March 2018 by Azad, T., Janse van Rensburg, H. J., et al.
Fig.5.G
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VEGFR regulates angiogenesis and tumor VM through YAP/TAZ in vitro. a Transient knockdown of YAP and/or TAZ in MCF10A overexpressing VEGFR2 decreases expression of ANG-2 and CYR61. Western blotting exposures indicate relative expression of YAP and TAZ. b, c VEGFR2 overexpression and VEGF treatmen...
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VEGFR regulates angiogenesis and tumor VM through YAP/TAZ in vitro. a Transient knockdown of YAP and/or TAZ in MCF10A overexpressing VEGFR2 decreases expression of ANG-2 and CYR61. Western blotting exposures indicate relative expression of YAP and TAZ. b, c VEGFR2 overexpression and VEGF treatment increases tube formation by MCF10A through YAP/TAZ. YAP and/or TAZ were transiently knocked down by siRNA in MCF10A stably overexpressing VEGFR2 and subjected to tube-formation assay on Matrigel 48 h after transfection alongside wild type MCF10A. For some conditions, cells were stimulated with 100 ng ml−1 VEGF or were treated with 100 nM verteporfin for the duration of the tube formation assay. Representative images are shown in b. Scale bar denotes 200 μm. Total tube formation was quantified in c (n = 3). *p < 0.05 in two-sample unpaired t-test. d–l YAP/TAZ are critical for VEGF-induced angiogenesis in HUVEC (d–f) and BOEC (g–i), as well as for vasculogenic mimicry in MDA-MB231 (j–l). d, g, j YAP and/or TAZ were transiently knocked down by siRNA in each cell line which reduced ANG-2 and CYR61 expression. Western blotting exposures indicate relative expression of YAP and TAZ for each cell line. e, h, k Representative images of tube formation on Matrigel assessed 48 h after transfection. For some conditions, cells were stimulated with 100 ng ml−1 VEGF or were treated with 100 nM verteporfin for the duration of the tube formation assay. Scale bar denotes 200 μm. Total tube formation was quantified in f, i, l (n = 3). *p < 0.05 in two-sample unpaired t-test. m, n Exogenous CYR61 and ANG-2 can partially rescue tube formation in YAP/TAZ knockdown HUVEC. YAP and/or TAZ were transiently knocked down by siRNA in HUVEC. Tube formation on Matrigel was assessed 48 h after transfection. For some conditions, cells were stimulated with 100 ng ml−1 VEGF, 200 ng ml−1 CYR61, and/or 200 ng ml−1ANG-2 for the duration of the tube formation assay. Scale bar denotes 200 μm. Representative images are shown in m and total tube formation was quantified in n. (n = 3). All data are represented as mean ± SD
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Collected and cropped from Nature Communications by CiteAb, provided under a CC-BY license
Image 1 of 32
In Oncotarget on 7 November 2017 by Shigeeda, W., Shibazaki, M., et al.
Fig.1.C
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Homo sapiens (Human)
RHAMM expression is regulated by YAP1/TAZ in MPM cells(A) Protein and (B) mRNA expression profiles of MPM cell lines. Numbers shown in (A) represent values relative to RHAMM/α-tubulin in Y-MESO-27 cells as a reference. (C) Protein and (D) mRNA expression profiles after single knockdown of YAP1, T...
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RHAMM expression is regulated by YAP1/TAZ in MPM cells(A) Protein and (B) mRNA expression profiles of MPM cell lines. Numbers shown in (A) represent values relative to RHAMM/α-tubulin in Y-MESO-27 cells as a reference. (C) Protein and (D) mRNA expression profiles after single knockdown of YAP1, TAZ, and RHAMM or concomitant knockdown of YAP1 and TAZ in MPM cell lines. Numbers shown in (C) represent values relative to RHAMM/α-tubulin in control siRNA-treated cells as a reference. (E) Protein and (F) mRNA expression profiles after forced expression of YAP1-5SA in MPM cell lines. “S” and “L” in (E) indicate “short” and “long” exposure, respectively. # and ## indicate Flag-tagged YAP1 5SA and endogenous YAP1, respectively. Bars in graphs indicate mean ± SEM of three independent experiments. Welch’s t test was used for statistical analysis (*P < 0.05, **P < 0.01).
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Collected and cropped from Oncotarget by CiteAb, provided under a CC-BY license
Image 1 of 32