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Fig.4.E showing Western Blotting in a Homo sapiens (Human) sample from the publication: Uvaol attenuates TGF-β1-induced epithelial-mesenchymal transition in human alveolar epithelial cells by modulating expression and membrane localization of β-catenin.

Fig.5.A showing Western Blotting from the publication: Liver X Receptors Enhance Epithelial to Mesenchymal Transition in Metastatic Prostate Cancer Cells.

Fig.1.B showing Western Blotting from the publication: Liver X Receptors Enhance Epithelial to Mesenchymal Transition in Metastatic Prostate Cancer Cells.

Fig.1.A showing Western Blotting from the publication: AGR2-mediated unconventional secretion of 14-3-3ε and α-actinin-4, responsive to ER stress and autophagy, drives chemotaxis in canine mammary tumor cells.

Fig.2.G showing Immunocytochemistry-immunofluorescence from the publication: Tumor-Extrinsic Axl Expression Shapes an Inflammatory Microenvironment Independent of Tumor Cell Promoting Axl Signaling in Hepatocellular Carcinoma.

Fig.4.A showing Immunocytochemistry-immunofluorescence in a Homo sapiens (Human) sample from the publication: Protective Effect of Limosilactobacillus fermentum ME-3 against the Increase in Paracellular Permeability Induced by Chemotherapy or Inflammatory Conditions in Caco-2 Cell Models.

Fig.2.B showing Immunocytochemistry-immunofluorescence from the publication: GJB4 variants linked to skin disease exhibit a trafficking deficiency en route to gap junction formation that can be restored by co-expression of select connexins.

Fig.7.F showing Western Blotting from the publication: Ladinin 1 Shortens Survival via Promoting Proliferation and Enhancing Invasiveness in Lung Adenocarcinoma.

Fig.1.B showing Western Blotting in a Homo sapiens (Human) sample from the publication: Downregulation of protein kinase C gamma reduces epithelial property and enhances malignant phenotypes in colorectal cancer cells.

Fig.6.C showing Western Blotting from the publication: Long noncoding RNA uc.230/CUG-binding protein 1 axis sustains intestinal epithelial homeostasis and response to tissue injury.

Fig.2.E showing Western Blotting from the publication: Pro-Apoptotic and Anti-Invasive Properties Underscore the Tumor-Suppressing Impact of Myoglobin on a Subset of Human Breast Cancer Cells.

Fig.5.D showing Western Blotting from the publication: Impaired fatty acid metabolism perpetuates lipotoxicity along the transition to chronic kidney injury.

Fig.7.C showing Western Blotting from the publication: Downregulated ADAMTS1 Incorporating A2M Contributes to Tumorigenesis and Alters Tumor Immune Microenvironment in Lung Adenocarcinoma.

Fig.3.D showing Western Blotting from the publication: SHC4 promotes tumor proliferation and metastasis by activating STAT3 signaling in hepatocellular carcinoma.

Fig.3.E showing Western Blotting from the publication: SHC4 promotes tumor proliferation and metastasis by activating STAT3 signaling in hepatocellular carcinoma.

Fig.5.E showing Western Blotting from the publication: SHC4 promotes tumor proliferation and metastasis by activating STAT3 signaling in hepatocellular carcinoma.

Fig.5.F showing Western Blotting from the publication: SHC4 promotes tumor proliferation and metastasis by activating STAT3 signaling in hepatocellular carcinoma.

Fig.6.F showing Western Blotting from the publication: SHC4 promotes tumor proliferation and metastasis by activating STAT3 signaling in hepatocellular carcinoma.

Fig.1.G showing Western Blotting in a Mus musculus (House mouse) sample from the publication: SYNCRIP Modulates the Epithelial-Mesenchymal Transition in Hepatocytes and HCC Cells.

Fig.4.D showing Western Blotting in a Homo sapiens (Human) sample from the publication: Hypoxia and heat stress affect epithelial integrity in a Caco-2/HT-29 co-culture.

Fig.3.E showing Immunocytochemistry-immunofluorescence from the publication: EPHA2-dependent outcompetition of KRASG12D mutant cells by wild-type neighbors in the adult pancreas.

Fig.3.D showing Western Blotting from the publication: Ubiquitin Conjugating Enzyme E2 H (UBE2H) Is Linked to Poor Outcomes and Metastasis in Lung Adenocarcinoma.

Fig.2.F showing Western Blotting from the publication: C3G downregulation induces the acquisition of a mesenchymal phenotype that enhances aggressiveness of glioblastoma cells.

Fig.1.C showing Western Blotting in a Homo sapiens (Human) sample from the publication: TGF-β-induced cell motility requires downregulation of ARHGAPs to sustain Rac1 activity.

Fig.4.E showing Western Blotting in a Homo sapiens (Human) sample from the publication: TGF-β-induced cell motility requires downregulation of ARHGAPs to sustain Rac1 activity.

146 images found

Defining enteric bacterial pathogenesis using organoids: Citrobacter rodentium uses EspC, an atypical mucinolytic protease, to penetrate mouse colonic mucus.

In Gut Microbes on 1 December 2025 by Chen, Y., Gilliland, A., et al.

Enteric bacterial pathogens pose significant threats to human health; however, the mechanisms by which they infect the mammalian gut in the face of daunting host defenses remain to be fully defined. For the attaching and effacing (A/E) bacterial family member and murine pathogen Citrobacter rodentium, its virulence strategy appears to involve penetration of the colonic mucus barrier to reach the underlying epithelium. To better define these interactions, we grew colonoids under air-liquid interface (ALI) conditions, producing a thick mucus layer that mimicked in vivo mucus composition and glycosylation. C. rodentium's penetration of ALI-derived mucus was dramatically enhanced upon exposure to sialic acid, in concert with the secretion of two serine protease autotransporter of Enterobacteriaceae (SPATE) proteins, Pic and EspC. Despite Pic being a class II SPATE, and already recognized as a mucinase, it was EspC, a class I SPATE family member, that degraded ALI-derived mucus, despite class I SPATEs not previously shown to possess mucinase activity. Confirming this finding, E. coli DH5α carrying a plasmid that expresses C. rodentium-derived EspC was able to degrade the mucus. Moreover, recombinant EspC alone also displayed mucinolytic activity in a dose-dependent manner. Collectively, our results reveal the utility of ALI-derived mucus in modeling microbe-host interactions at the intestinal mucosal surface, as well as identify EspC as an atypical class I SPATE that shows significant mucinolytic activity toward ALI-derived mucus.

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PCK2 promotes invasion and epithelial-to-mesenchymal transition in triple-negative breast cancer by promoting TGF-β/SMAD3 signaling through inhibiting TRIM67-mediated SMAD3 ubiquitination.

In Cancer Biology & Therapy on 1 December 2025 by Chang, T. M., Fang, W. Y., et al.

PCK2, which encodes mitochondrial phosphoenolpyruvate carboxykinase (PEPCK-M), is upregulated in various cancers. We demonstrated high expression of PEPCK-M in approximately half of triple-negative breast cancers (TNBCs) previously. TNBC is associated with an aggressive phenotype and a high metastasis rate. In this study, we investigated the role of PCK2 in TNBC. PCK2 knockdown suppressed proliferation and mTOR signaling in TNBC cells. In addition, cell invasion/migration ability and the expression of epithelial-to-mesenchymal transition (EMT) markers were positively correlated with PCK2 expression in TNBC cells via regulation of transforming growth factor-β (TGF-β)/SMAD3 signaling. SMAD3 was positively regulated by PCK2 in TNBC cells. Knockdown of SMAD3 in PCK2-overexpressing TNBC cells reduced the expression levels of EMT markers, Snail and Slug, and suppressed cell invasion/migration. In addition, PCK2 knockdown attenuated the stimulatory effect of TGF-β on SMAD3 phosphorylation in TNBC cells. PEPCK-M promotes the protein and mRNA expression of SMAD3 via competitive binding to tripartite motif-containing 67 (TRIM67), an E3 ubiquitin ligase, to reduce SMAD3 ubiquitination, which leads to promoting nuclear translocation of SMAD3 and autoregulation of SMAD3 transcription. Moreover, high PCK2 mRNA expression was significantly associated with poor survival in TNBC patients. In conclusion, our study revealed for the first time that PCK2 activates TGF-β/SMAD3 signaling by regulating the expression and phosphorylation of SMAD3 by inhibiting TRIM67-mediated SMAD3 ubiquitination and promoting the stimulatory effect of TGF-β to promote TNBC invasion. The regulatory effect of PCK2 on mTOR and TGF-β/SMAD3 signaling suggests that PCK2 is a potential therapeutic target for suppressing TNBC progression.

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Activation of Human FPR2 with AT-RvD1 Resolves Acute Sialadenitis in Vivo.

In Inflammation on 4 June 2025 by Nam, K., Dos Santos, H. T., et al.

Previous studies demonstrated that activation of the mouse G protein-coupled formyl peptide receptor 2 (mFpr2) with aspirin-triggered resolvin D1 (AT-RvD1) blocks pro-inflammatory cytokine signaling while promoting salivary gland (SG) epithelial integrity both in vitro and in vivo. In addition, mice lacking Fpr2 display alterations of SG innate and adaptive immunity. Taken together, these results indicate that Fpr2 activation with AT-RvD1 restores saliva secretion and regulates SG immunity in mice. To demonstrate the value of AT-RvD1 for use in human SG, however, we need to extend the findings above in the direction of clinical use. To this end, the current study investigated whether treatment with AT-RvD1 reduces SG inflammation and restores saliva secretion in an acute sialadenitis mouse model expressing the human formyl peptide receptor 2 (hFPR2) protein. Results indicate that mice carrying the hFPR2 and treated with lipopolysaccharide (LPS) display acute sialadenitis-like features as shown by increased levels of proliferating inflammatory cells, loss of epithelial integrity and reduced saliva secretion. In contrast, when these mice are treated with AT-RvD1, the sialadenitis-like features are drastically reduced as evidenced by a significant decrease in proliferating inflammatory cells as well as restoration of saliva secretion to levels comparable to phosphate buffered saline (PBS)-treated healthy controls. Finally, changes observed in mice carrying the hFPR2 and treated with LPS and AT-RvD1 were comparable to those observed in wild-type mice carrying the mFpr2. Together, these results demonstrate that activation of hFPR2 with AT-RvD1 resolves acute sialadenitis in vivo.
© 2025. The Author(s).

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Identification of FLYWCH1 as a regulator of platinum-resistance in epithelial ovarian cancer.

In NAR Cancer on 1 June 2025 by Bauer, T. L., Rohm, H., et al.

Platinum-based combination chemotherapy remains the backbone of first-line treatment for patients with advanced epithelial ovarian cancer (EOC). While most patients initially respond well to the treatment, patients with relapse ultimately develop platinum resistance. This study identified FLYWCH-type zinc finger-containing protein 1 (FLYWCH1) as an important regulator in the resistance development process. We showed that the loss of FLYWCH1 promotes platinum resistance in EOC cells, and the low FLYWCH1 expression is correlated with poor prognosis of EOC patients. In platinum-sensitive cells, FLYWCH1 colocalizes with H3K9me3, but this association is significantly reduced when cells acquire resistance. The suppression of FLYWCH1 induces gene expression changes resulting in the deregulation of pathways associated with resistance. In line with its connection to H3K9me3, FLYWCH1 induces gene silencing in a synthetic reporter assay and the suppression of FLYWCH1 alters H3K9me3 at promoter regions and repeat elements. The loss of FLYWCH1 leads to the derepression of LTR and Alu repeats, thereby increasing transcriptional plasticity and driving the resistance development process. Our data highlight the importance of FLYWCH1 in chromatin biology and acquisition of platinum resistance through transcriptional plasticity and propose FLYWCH1 as a potential biomarker for predicting treatment responses in EOC patients.
© The Author(s) 2025. Published by Oxford University Press on behalf of NAR Cancer.

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Bat organoids reveal antiviral responses at epithelial surfaces.

In Nature Immunology on 1 June 2025 by Kellner, M. J., Monteil, V., et al.

Bats can host viruses of pandemic concern without developing disease. The mechanisms underlying their exceptional resilience to viral infections are largely unresolved, necessitating the development of physiologically relevant and genetically tractable research models. Here, we developed respiratory and intestinal organoids that recapitulated the cellular diversity of the in vivo epithelium present in Rousettus aegyptiacus, the natural reservoir for the highly pathogenic Marburg virus (MARV). In contrast to human counterparts, bat organoids and mucosal tissue exhibited elevated constitutive expression of innate immune effectors, including type I interferon-ε (IFNε) and IFN-stimulated genes (ISGs). Upon infection with diverse zoonotic viruses, including MARV, bat organoids strongly induced type I and III IFN responses, which conferred robust antiviral protection. Type III IFNλ3 additionally displayed virus-independent self-amplification, acting as an ISG to enhance antiviral immunity. Our organoid platform reveals key features of bat epithelial antiviral immunity that may inform therapeutic strategies for viral disease resilience.
© 2025. The Author(s).

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AGR2-mediated unconventional secretion of 14-3-3ε and α-actinin-4, responsive to ER stress and autophagy, drives chemotaxis in canine mammary tumor cells.

In Cell Mol Biol Lett on 31 May 2024 by Yuan, S. H., Wu, C. C., et al.

Fig.1.A

Fig.1.A showing Western Blotting from the publication: AGR2-mediated unconventional secretion of 14-3-3ε and α-actinin-4, responsive to ER stress and autophagy, drives chemotaxis in canine mammary tumor cells.

Ectopic expression of AGR2 modulated extracellular milieu of CMT cells, promoting CMT cell chemotaxis. A Characterization of CMT cell lines. Expression of AGR2, E-cadherin, vimentin, HER-2, or ER-⍺ in individual cell lines was analyzed by immunoblotting with specific antibodies. CMT-U27 (B) and C... more

Ectopic expression of AGR2 modulated extracellular milieu of CMT cells, promoting CMT cell chemotaxis. A Characterization of CMT cell lines. Expression of AGR2, E-cadherin, vimentin, HER-2, or ER-⍺ in individual cell lines was analyzed by immunoblotting with specific antibodies. CMT-U27 (B) and CF41.Mg (C) were transfected with pcDNA3.1-myc.His-AGR2 or the mock vector and grown in 2% FBS-containing RPMI and DMEM, respectively, for 24 h. Whole-cell lysates (WCL) of the transfectants were analyzed by immunoblotting to confirm the expression of Myc-tagged AGR2. Conditioned media (CM) of the transfectants were collected and placed in the bottom well for a transwell migration assay, in which the responding cells were seeded in the top insert and stained with Hoechst during a 16-h incubation. Cells in the insert were fixed for image acquisition using an epifluorescence microscope with a 10 × objective. D, F The number of migrated cells was counted and presented as the mean + SD of three independent experiments. E, G Representative images of migrated cells per field were shown. H Myc-Trap-based precipitation conducted the depletion of AGR2 in the CM of AGR2-expressing CMT-U27. Expression of AGR2 in WCL or CM and Myc-Trap-precipitated AGR2 were verified by immunoblotting. I As described above, the AGR2-depleted CM (denoted deAGR2) or the untreated control was placed in the bottom well for a transwell migration assay. J, K Fresh 2% FBS-containing media supplemented with or without rcAGR2 (800 ng/mL) were placed in the bottom well for a transwell migration assay. For C, F, I–K, statistical significance was determined by a two-tailed unpaired t-test. *p < 0.05; **p < 0.01; ****p < 0.0001 less

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Pro-Apoptotic and Anti-Invasive Properties Underscore the Tumor-Suppressing Impact of Myoglobin on a Subset of Human Breast Cancer Cells.

In Int J Mol Sci on 29 September 2022 by Aboouf, M. A., Armbruster, J., et al.

Fig.2.E

Fig.2.E showing Western Blotting from the publication: Pro-Apoptotic and Anti-Invasive Properties Underscore the Tumor-Suppressing Impact of Myoglobin on a Subset of Human Breast Cancer Cells.

Loss of myoglobin increases the migratory capacity of hypoxic MCF7 cancer cells (A) Representative images of wound healing assay for MCF7 clones grown at 0.2% O2 hypoxia (Hx) in serum reduced medium for 48 h. A scratch was initiated into a monolayer of each cell line and cells were incubated in n... more

Loss of myoglobin increases the migratory capacity of hypoxic MCF7 cancer cells (A) Representative images of wound healing assay for MCF7 clones grown at 0.2% O2 hypoxia (Hx) in serum reduced medium for 48 h. A scratch was initiated into a monolayer of each cell line and cells were incubated in normoxia (Nx) or 0.2% O2 Hx. The % of the scratch area was measured and calculated after 24 and 48 h (n = 3). (B) Scatter plot of % scratch area (normalized to 0 h) against the time of incubation. MCF7 (left panels) and SKBR3 (right panels) wildtype (MB-WT, white) and MB knockout (MB-KO, red) clones (n = 3). (C) Representative images of Transwell invasion assay showing only invading wildtype and MB-KO MCF7 cells through a layer of Matrigel after culturing at 0.2% O2 Hx in Transwell for 48 h followed by fixation/staining with 0.6% glutaraldehyde and crystal violet solution. Magnification is 5x. (D) Quantitative representation of % of crystal violet signal area of invading cells of MB-WT (white) and MB-KO (red) of MCF7. Five random images from each well were captured and analyzed by ImageJ and the average of the signal area was calculated (n = 3). (E) Representative Western blotting image (n = 3) of whole tissue lysate of MB-WT and MB-KO clones of MCF7 cells cultured at 0.2% O2 Hx for 72 h and stained for Smad2/3 (60 and 52 kDa, respectively phospho-Smad2 (60 kDa phospho-Smad 1/5/9 (60 kDa), E-Cadherin (120 kDa), PI3K-p85 (85 kDa), and β-actin (42 kDa) used as the loading control. (F) Band intensity of Smad2, phospho-Smad2, phospho-Smad 1/5/9, E-Cadherin, and PI3K-p85 proteins after Western blotting, from MB-WT (white bars) and MB-KO (red bars) cells at 0.2% O2 Hx, was quantified using MCID Analysis 7.0 and normalized to β-actin (n = 3). (G) Relative mRNA expression levels of genes that regulate epithelial to mesenchymal transition: TWIST, SNAIL, SLUG, FN1, ZEB1, MMP3, PI3K CA, and Pi3K CB, quantified by qPCR and normalized to β-actin (ACTB) mRNA expression levels, from MB-WT (white) and MB-KO (red) clones of MCF7 cells cultured at 0.2% O2 Hx for 72 h. (n = 3 per group). Data are presented as mean and standard error of mean and analyzed by Student’s t-test * p < 0.05; ** p < 0.01, *** p < 0.001. less

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SYNCRIP Modulates the Epithelial-Mesenchymal Transition in Hepatocytes and HCC Cells.

In Int J Mol Sci on 14 January 2022 by Riccioni, V., Trionfetti, F., et al.

Fig.1.G

Fig.1.G showing Western Blotting in a Mus musculus (House mouse) sample from the publication: SYNCRIP Modulates the Epithelial-Mesenchymal Transition in Hepatocytes and HCC Cells.

SYNCRIP is involved in TGFβ-induced EMT of hepatocytes. (A) Western blot analysis of intracellular levels of SYNCRIP in hepatocytes (3A) treated or not for 24 h with TGFβ. GAPDH was used as loading control. All the experiments were performed three times and images are representative of one indica... more

SYNCRIP is involved in TGFβ-induced EMT of hepatocytes. (A) Western blot analysis of intracellular levels of SYNCRIP in hepatocytes (3A) treated or not for 24 h with TGFβ. GAPDH was used as loading control. All the experiments were performed three times and images are representative of one indicative experiment of the independent ones. The densitometry analysis (right panel) was conducted by using Image J software. Data are reported as means ± SD. Statistically significant differences are reported (* p < 0.05). (B) qRT-PCR analysis of Syncrip intracellular levels in hepatocytes stably silenced for SYNCRIP (3A shSYN) compared to cells with the empty vector (3A shCTR). The values are calculated via the 2 (−ΔCt) method, normalized to the 18S ribosomal RNA levels and shown as mean ± SD. Statistically significant differences are reported for four independent experiments (* p < 0.05). (C) Western blot analysis of SYNCRIP and GAPDH as loading control. The image is representative of four independent experiments. The densitometry analysis (right panel) was conducted by using the Image J software. Data are means ± SD of four independent experiments. Statistically significant differences are reported (*** p < 0.001). (D) Phase contrast micrographs of 3A shSYN cells or 3A shCTR, treated or not for 24 h with TGFβ as indicated. (E) Immunofluorescence assays for epithelial (ZO-1 and E-Cadherin) and mesenchymal (SNAIL and FIBRONECTIN) markers in TGFβ-treated or untreated 3A shCTR and 3A shSYN cells. Nuclei were stained with Hoechst (blue). Images are representative of three independent experiments. Scale bar is indicated. (F) qRT-PCR analysis for the indicated mesenchymal (Snail and Zeb2) and epithelial (E-cadherin and HNF1α) markers in TGFβ-treated or untreated 3A shCTR and 3A shSYN cells. The values are calculated by the 2 (−ΔCt) method, normalized to the 18S ribosomal RNA levels and shown as mean ± SD. Statistically significant differences are reported for five independent experiments (* p < 0.05; ** p < 0.01; ns = no significance). (G) Western blot analysis of intracellular levels of the indicated epithelial and mesenchymal markers in TGFβ-treated or untreated 3A shCTR and 3A shSYN cells. The image is representative of four independent experiments. GAPDH was used as loading control. The densitometry analysis (right panel) was conducted by using the Image J software. Data are means ± SD of four independent experiments. Statistically significant differences are reported (* p < 0.05; ** p < 0.01; ns = no significance). less

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C3G downregulation induces the acquisition of a mesenchymal phenotype that enhances aggressiveness of glioblastoma cells.

In Cell Death Dis on 6 April 2021 by Manzano, S., Gutiérrez-Uzquiza, A., et al.

Fig.2.F

Fig.2.F showing Western Blotting from the publication: C3G downregulation induces the acquisition of a mesenchymal phenotype that enhances aggressiveness of glioblastoma cells.

C3G downregulation enhances invasion of GBM cells promoting the expression of mesenchymal markers.Non-silenced or C3G-silenced (shC3G) U87 and 12Ф12D cells, and U87 cells with control shRNA (NTC) have been used. A and B Invasion through Matrigel of U87 and 12Ф12D cells (respectively) using FBS as... more

C3G downregulation enhances invasion of GBM cells promoting the expression of mesenchymal markers.Non-silenced or C3G-silenced (shC3G) U87 and 12Ф12D cells, and U87 cells with control shRNA (NTC) have been used. A and B Invasion through Matrigel of U87 and 12Ф12D cells (respectively) using FBS as chemoattractant (10%). Left panels, representative images of invading cells; right panels, histograms showing the mean value ± S.E.M. of the number of invading cells (n = 3). C–E Adhesion analysis at 15 or 30 min in U87 and 12Ф12D cells. Top panels, phase contrast microscopy representative images of adhered U87 cells on uncoated dishes (no ECM) or coated with Matrigel; lower panels, histograms showing mean value ± S.E.M. of the percentage of adhered cells referred to non-silenced cells at 15 min (n = 3). *p ≤ 0.05, **p ≤ 0.01 and ***p ≤ 0.001, compared as indicated. Scale bars (A–E): 100 µm. F Representative western-blot analyses of Vimentin and E-cadherin levels normalized to β-actin in U87 (upper panel) and 12Ф12D cells (lower panel) treated as indicated (n = 3). GTWIST1 and ZEB2 mRNA levels in U87 cells maintained either in the presence (+) or absence (−) of serum. Histograms represent RQ mean value ± S.E.M. referred to parental cells maintained with 10% FBS (n = 2–4). *p ≤ 0.05, compared as indicated. less

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TGF-β-induced cell motility requires downregulation of ARHGAPs to sustain Rac1 activity.

In J Biol Chem on 21 March 2021 by Motizuki, M., Koinuma, D., et al.

Fig.1.C

Fig.1.C showing Western Blotting in a Homo sapiens (Human) sample from the publication: TGF-β-induced cell motility requires downregulation of ARHGAPs to sustain Rac1 activity.

EMT-associated cellular responses that are dependent on Smad3. A, SMAD3 knockout A549 cells (A549-S3KO) were prepared using CRISPR/Cas9-mediated genome editing. A549-S3KO cells were infected with lentivirus carrying SMAD3 cDNA. Expression and TGF-β-induced phosphorylation of Smad3 were verified b... more

EMT-associated cellular responses that are dependent on Smad3. A, SMAD3 knockout A549 cells (A549-S3KO) were prepared using CRISPR/Cas9-mediated genome editing. A549-S3KO cells were infected with lentivirus carrying SMAD3 cDNA. Expression and TGF-β-induced phosphorylation of Smad3 were verified by immunoblotting with the indicated antibodies; α-tubulin was used as a loading control. B–E, A549 cells, A549-S3KO cells, or those expressing wild-type Smad3 were incubated in either the presence or absence of 1 ng/ml TGF-β1 for 24 h. B, light microscopic photographs. C, expression of E-cadherin was determined by immunoblotting with anti-E-cadherin; α-tubulin was used as a loading control. D, immunofluorescence detection of E-cadherin. E, formation of actin stress fibers in A549-S3KO cells expressing wild-type Smad3. F-actin was stained using Rhodamine-phalloidin. F, chamber migration assay or (G) wound healing assay of A549 cells, A549-S3KO cells, or those expressing wild-type Smad3 in either the presence or absence of 1 ng/ml TGF-β1. Quantification is shown in the right. H, to evaluate growth rates of A549 cells, A549-S3KO cells, or those expressing wild-type Smad3 in either the presence or absence of 1 ng/ml TGF-β1, cell numbers were counted. TGF-β was added at day1. Scale bars: 10 μm (B and D–F) and 200 μm (G). Error bars represent SD (n = 5 for F, G). p values were determined by Student's t-test. ∗p < 0.01. One representative result from two independent experiments is shown (F–H). less

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