Fig. 6.

miR-222 regulates p27kip1 and PUMA expression to induce cell death in SEB-activated splenocytes. Splenocytes were transfected with control (mock) or miRNA mimic and inhibitor for miRNA-222 in the presence of SEB (1 μg/ml). Eighteen hours later, cells were treated with DIM (25 μM) or vehicle for 6 hours. (A) Total RNA was isolated from cells and analyzed for miR-222 expression by qPCR. (B) Cells were collected and stained with AnnexinV/PI staining to determine apoptosis and then quantified for cell death. (C) Relative expression level of gene targets p27kip1 and PUMA by quantitative PCR. Data are presented as means ± S.E.M. (n = 3 per group). *, **, and *** represent significant difference with P values < 0.05, < 0.01, and < 0.001, respectively.