Figure 1.

Impact of resveratrol on TNF-β-induced epithelial-to-mesenchymal transition (EMT) and EMT-related proteins in CRC cells (HCT116, RKO, SW480). CRC cells in monolayer culture were either left untreated (Co) or treated with 5µM resveratrol (Res), 10 ng/mL TNF-β, or 10ng/mL TNF-α for 12 h, or cells were pre-treated with resveratrol (5µM) for 4 h, followed by co-treatment with 10ng/mL TNF-β or 10ng/mL TNF-α for 12 h. (A) Cell phenotype changes associated with EMT, as demonstrated in phase contrast images. Cell colonies (arrows). Scale bar = 100 μm. (B) Immunoblotting of cell lysates was performed for E-cadherin, vimentin, and slug. The housekeeping protein β-actin served as an internal loading control. (C) Immunolabeling was done with primary antibodies for E-cadherin, vimentin, and slug, followed by incubation with rhodamine-coupled secondary antibodies. All experiments were performed at least in triplicate and quantification of positively labelled cells was done by counting 600–800 cells from 10 microscopic fields. All data were compared to the control and statistically significant values with p < 0.05 were designated by (*) and p < 0.01 were designed by (**). Original magnification, 600×. Scale bar = 30 nm.