E47-silencing in MDCK-EGFP-E47 cells induces a MET process, downregulation of Id1 and reduction of tumour growth.

<p>(<b>A</b>) Phase contrast images (a-e) and immunofluorescence images (f–y) of the indicated cell lines showing localization of EGFP (f,g), EGFP-E47 (h–j), Ecadherin (k–o), beta-catenin (p–t) and F-actin organization (u–y). Nuclei were stained with DAPI (k–y). Black bar, 60 um (a–e); white bars, 25 um (f–y). (<b>B</b>) Western blot analyses of epithelial markers and Id1 in the indicated cell lines; alpha-tubulin was used as loading control. Two (#N, #P) and five independent clones (#N, #M, #L, #O, #P) generated after stable expression of shEGFP into MDCK-EGFP-E47 (clone #6) cells are shown in <b>A</b> and <b>B</b>, respectively. (<b>C</b>) The tumorigenic potential of MDCK-EGFP-E47#6 and MDCK-EGFP-E47-shEGFP#N and #P cells was analysed by subcutaneous injection into nude mice. ANOVA analysis: *p<0.05; **p<0.01; ***p<0.001. (<b>D</b>) Histology (upper panels) and immunohistochemical analysis for E-cadherin (lower panels) of tumours induced by MDCK-EGFP-E47#6 (left) and MDCK-EGFP-E47-shEGFP#N cells (right). Amplification x20. Insets show amplified fields at x40.