Nrf2-silencing represses the increase in the levels of antioxidant enzymes, MMP2 and VEGF induced by EtOH treatment. (A,B) HCT116 cells (1 × 105) were transiently transfected with a nonspecific siRNA (siControl) or with Nrf2 specific siRNA pool (siNrf2) and, 24 h after transfection, cells were treated with 300 mM EtOH for additional 24 h or 48 h. (A) The levels of HO-1, MnSOD, MMP2, and VEGF were analyzed by Western blotting after 24 h of transfection. The successful of Nrf2-silencing was verified by measuring the level of Nrf2 in transfected cells. Quantitative estimations of the protein levels were determined by densitometry measurements of western blotting from three independent experiments after normalization with β-actin (*) p < 0.05 and (**) p < 0.01 compared to the untreated sample. (#) p < 0.05, (##) p < 0.01. (B) Effects of Nrf2-silencing on the viability of HCT116 cells evaluated after 48 h of EtOH treatment by MTT assay and expressed as the percentage of the viable siControl cells in untreated cultures. Values are the means of three independent experiments ± S.E. (#) p < 0.05 between the two groups.