Fig. 7

Effect of knocking down p38γ MAPK on alcohol-induced tumor promotion. a The expression of p38γ and p38α MAPK in MCF7-ErbB2 cells stably expressing control shRNA (Consh) or p38γ shRNA (p38γsh) was determined by immunoblotting. b The relative levels of p38γ in Consh and p38γsh-treated cells were quantified and expressed relative to Consh-treated group. c and d MCF7-ErbB2 and BT474 cells stably expressing control Consh or p38γsh were exposed to alcohol (0 or 100 mg/dl) for 10 days. After alcohol exposure, 1000 cells/well were cultured on ultra-low attachment plates for 10 days. The number of mammospheres was counted and calculated relative to the control groups treated with Consh. e After alcohol exposure for 10 days, the CSC population (ALDEFLUOR positive cells) in MCF7-ErbB2 cells stably expressing control shRNA (Consh) and p38γ shRNA (p38γsh) was determined by ALDEFLUOR assay followed by flow cytometry as described in the Materials and Methods. f and g After alcohol exposure for 10 days, MCF7-ErbB2 cells stably expressing control shRNA (Consh) or p38γ shRNA (p38γsh) were assayed for migration and invasion as described in the Materials and Methods. Each data point was the mean ± SEM of three independent experiments and expressed relative to control groups. *denotes significant difference between non-alcohol-treated groups (p38γsh-treated controls vs Consh-treated controls). #denotes significant difference from respective alcohol-exposed cells expressing p38γsh