Fig. 3: Enforced overexpression of miR-22 repressed 182^R-6 cell proliferation, induced apoptosis, and caused cell cycle S-phase arrest via overexpression of p21 and p27.

a 182^R-6 cells grown to 80% confluency in a DMEM/F12 medium supplemented with 0.1 μM fulvestrant were transfected with either 40 nM miR-22 mimic or AllStars negative control siRNA. At 72 h after transfection, total RNA was isolated and subjected to qRT-PCR analysis using a primer set of hsa-miR-22; RNU6-2 was used as a reference gene to normalize miR-22 expression. b At  24 h after transfection, the 182^R-6 cells were reseeded in 96-well plates, and the MTT assay was performed as described in the “Methods” section. c At 72 h after transfection, the 182^R-6 cells were harvested, and cell cycle analysis was performed as described in the “Methods” section. d At 72 h after transfection, the 182^R-6 cells were harvested, and apoptosis analysis was performed as described in the “Methods” section. e At 72 h after transfection, whole cell lysates were prepared and subjected to western blot analysis using the indicated antibodies, as described in the “Methods” section. Actin served as a loading control. * indicates p < 0.05