Smartpool short inhibitory RNA (siRNA) for human Snail (Snail1, SNAI1) was purchased from Dharmacon (Dharmacon, Inc., Lafayette, CO). Caco-2 cells grown to 60% confluence in 6-well plates were transfected with 100nM Snail siRNA or control siRNA (Santa Cruz Biotechnology), using Lipofectamine (Invitrogen) and Optimem (Invitrogen) media (Forsyth et al., 2007). At 2 days post siRNA transfection cells were treated with alcohol (0.2%, 43 mM) for 4 additional days and then tested for expression of Snail and vimentin proteins by western blotting. Blot shown is representative from 3 experiments while densitometry data are means ± SE from all three experiments.
