p130Cas silencing promotes acquirement of epithelial traits in A17 cells. (A) Extracts from A17 cells expressing scramble (Ctr shRNA) or p130Cas shRNAs (p130Cas shRNA) treated (days: 4) or not (days: 0) with doxycycline were blotted with antibodies to p130Cas. Actin was used as loading control. (B) Representative images of one out of ten experiments performed in A17 cells as in (A) (a, b) and p130Cas silenced cells reconstituted with human full-length (hFL) p130Cas (p130Cas shRNA + hFL p130Cas) (c) cultured in presence of doxycycline for four days (20X magnification). Right panel: Quantification of length/width ratio on five distinct microscope fields in each condition (**P value <0.001). (C) Extracts from cells as in (B) were probed with antibodies to p130Cas, Snail, Slug, Twist, Vimentin, and normalized with Vinculin. (D) Left panels: quantification of E-cadherin mRNA by qRT-PCR in Control shRNA, p130Cas shRNA and in p130Cas silenced cells reconstituted with human full-length p130Cas (hFLCas) A17 cells (**P <0.001). Right panel: cell extracts were blotted with antibodies to p130Cas, and E-cadherin. Actin was used as loading control. (E) Representative images of E-cadherin immunofluorescence performed on cells as in (D) (100X magnification).