FIGURE 4.

TGFβ pathway is involved in the EMT phenotype in MCF10A retroviral cells. A, pan-TGFβ, total Smads2/3/4, and phospho-Smad2/3 protein levels were analyzed using Western blotting and showed that phospho-Smad2/3 is increased in UTR containing cell only (β-actin detected using the rabbit anti-actin antibody). B, immunohistochemistry of phospho-Smad2/3 complex in control cells and cells depleted of p63α and β (UTR) shows that only in UTR-containing cells is the Smad2/3 phosphorylated in the nucleus. C, phase-contrast images of MCF10A cells with SCR and UTR shRNA, with and without the addition of the TGFβ pathway inhibitor SB-431542, show a rescue of the epithelial cell phenotype. D, protein levels of total Smad2 and 3, phospho-Smad2 and 3, Slug, Snail, and Twist were analyzed by Western blotting, in SCR and UTR cells with and without TGFβ inhibitor and showed that inhibition reduced phospho-Smad2/3 levels as well as total levels of Slug and Snail. All experiments were carried out in triplicate, and three independent studies were performed.