E2A expression inhibits EMT in CRC cells. A, Knockdown of E2A induced EMT and MMP-9 expression. Left panel: Immunoblot analysis showed epithelial marker E-cadherin was decreased and mesenchymal markers vimentin and β-catenin were increased in shE2A-expressing SW480 clone, indicating induction of EMT. MMP-9 was also increased upon E2A downregulation, which demonstrated enhanced invasion ability. GAPDH was used as loading control. Right panel: Densitometric analysis of left panel normalized to GAPDH. B, Expression of E12 and E47 suppressed EMT and MMP-9 expression. Left penal: Immunoblot analysis of E-cadherin, vimentin, β-catenin, MMP-9 and GAPDH expression with or without transfection of E12 or E47. Right penal: Densitometric analysis of left normalized to GAPDH. Data in the histograms is expressed as the means ± SD from 3 separate experiments. C, Immunofluorescence analysis using anti- E-cadherin, vimentin, β-catenin antibodies as indicated. Nuclei were counterstained with 4′, 6-diamidino-2-phenylindole (DAPI). Magnification: 400×; Scale: 50 μm. *, P < 0.05; **, P < 0.01.