Fig. 4

TWIST1 regulates the expression of E-cadherin, vimentin, β-catenin, Bcl2 and Bax proteins in ESCC cell lines. Representative images of ICC staining (in brown) for (A, B) E-cadherin, (C, D) vimentin, (E, F) β-catenin, (G, H) Bcl2, (I, J) Bax in KYSE-30 (upper panel) or YM-1 (lower panel) cells expressing GFP + TWIST1 or GFP as a control. Nuclei are stained with hematoxylin. The blue color indicates negative immunostaining. K Bar charts showing the percentage of positive cells for E-cadherin, vimentin, β-catenin, Bcl2, and Bax proteins to compare the TWIST1-expressing cells with the control cells. Twenty randomly chosen fields was counted at 40X magnification (≥ 150 cells) for each sample and the values of positive cells (%) were averaged. * P < 0.01. Scale bar: 100 μm