FIGURE 8:

CUGBP1 and miR-222 synergistically repress CDK4 translation. (A) Changes in the levels of CDK4 protein expression after miR-222 overexpression with or without CUGBP1 silencing. After cells were cotransfected premiR-222 and control-siRNA (C-siRNA) or siRNA targeting CUGBP1 (siCUGBP1) for 48 h, CUGBP1 and CDK4 protein levels were examined. (B) CDK4 translation efficiency was measured by analysis of CDK4 CR luciferase reporter in cells described in (A). Values are the means ± SE of data from three separate experiments. *p < 0.05 compared with controls and cells transfected with control scrambled oligomer. +p < 0.05 compared with cells cotransfected with premiR-222 and C-siRNA. (C) Effects of reducing miR-222 alone or reducing both miR-222 and CUGBP1 on CDK4 protein expression in polyamine-deleted cells. After cells were treated with DFMO for 4 d, they were transfected with control oligomers, anti-miR-222 oligos, or anti-miR-222 oligos plus siCUGBP1; protein levels were measured 48 h after transfection. (D) CDK4 translation efficiency in cells described in (B).