Figure 6
Representative confocal images of double cytokeratin and vimentin (A), β-catenin (B) or E-cadherin (C) immunofluorescence corresponding to 2D endometrial epithelial monolayers isolated from Cre:ER+/−; SMAD2f/f; SMAD3f/f, plated and treated with tamoxifen (TAM) to induce Cre:ER-mediated deletion of SMAD2 and SMAD3 alleles or left untreated (NO TAM). Images were captured after 3 days of tamoxifen-induced deletion. Cells were counterstained with Hoechst to visualize nuclei. Scale bars: 25 µm. D. Western blot analysis of vimentin expression on 2D endometrial epithelial monolayers isolated from Cre:ER+/−; SMAD2f/f; SMAD3f/f. Cells were treated (TAM) or not (NO TAM) with tamoxifen to induce Cre:ER-mediated deletion of SMAD2 and SMAD3 alleles and then stimulated or not with TGF-β for 72 h. Membranes were also blotted with β-actin antibody to ensure deletion of these proteins. E. RT-qPCR relative quantification of Cdh2 and Twist1 mRNA expression of 2D endometrial epithelial monolayers isolated from Cre:ER+/−; SMAD2f/f; SMAD3f/f. Cells were plated and treated (TAM) or not (NO TAM) with tamoxifen to induce Cre:ER-mediated deletion of SMAD2 and SMAD3 alleles. Values are mean and error bars represent mean ± S.E.M. *P < 0.001 by one-way ANOVA.
