Figure 2.

Validation of selected gene targets identified in microarray analyses. A. RanBPM shRNA Hela (left panel) and HCT116 (right panel) cells were transfected with pCMV-RanBPM si-mt construct. Forty-eight hours post-transfection, whole cell extracts were prepared and analyzed by western blotting, alongside control and untransfected RanBPM shRNA extracts. Restoration of RanBPM expression was verified by hybridizing with a RanBPM antibody. B. cDNA from control shRNA, RanBPM shRNA, and RanBPM shRNA +RanBPM si-mt Hela (top panel) and HCT116 (bottom panel) was analyzed by RT-qPCR analyses using primers specific to the indicated genes. Fold gene expression was normalized to control shRNA cells. Genes appearing to the left of x-axis break are genes whose expression was responsive to restoration of RanBPM expression, while genes appearing to the right of axis break were not responsive to restored RanBPM expression. Data represents the mean of a minimum of three independent experiments with error bars indicating SE, * P < 0.05, ** P < 0.005.