Fig 7.

The role of cyclin D1 in MIC-1-associated survival of EPEC-infected intestinal epithelial cells. (A) HCT-8 intestinal epithelial cells were infected with E. coli K-12, wild-type EPEC, or EPEC ΔescN at a ratio of 50:1 (bacteria to cells) at each time point. Total proteins from the epithelial cellular lysate were subjected to Western blot analysis. (B) Stable cell lines (vector- and shMIC-1-transfected HCT-8 cells) were pretreated with 5 μM BAY 11–7082. Next, the cells were infected with EPEC at a ratio of 50:1 (bacteria to cells) for 12 h. Apoptosis was quantified by staining with propidium iodide (PI) and performing fluorescence-activated cell sorting analysis. All results are representative of three independent experiments. (C) Stable cell lines (vector- and shMIC-1-transfected HCT-8 cells) were pretreated with vehicle (dimethyl sulfoxide), 20 μM BAY11-7082, or 100 nM (5Z)-7-oxozeaenol and then infected with EPEC for 12 h. The total epithelial cellular lysates were subjected to real-time RT-PCR. (D) HCT-8 cells expressing either control or cyclin D1-specific shRNA were infected with EPEC at a ratio of 50:1 (bacteria to cell) for 12 h. Apoptosis was quantified by staining with propidium iodide (PI) and performing fluorescence-activated cell sorting analysis. Values are the means ± standard errors of the means (n = 3 to 6). Bars with different letters are significantly different (P < 0.05). Pairwise comparisons were made using SNK post hoc ANOVA.