Figure 5

PHLPP2 and FOXO1 are potential targets of miR-135a. A. Sequence of PHLPP2- or FOXO1-3′UTR miR-135a binding seed region, miR-135 and mutation of miR-135a (shown as miR-135a-mut). B. Upper: Western blotting analysis of PHLPP2 or FOXO1 in bladder cancer tissues, compared with normal bladder tissue. α-Tubulin was used as a loading control. Lower: Real-time PCR analysis of miR-135a in bladder cancer tissues, and the correlation between miR-135a and PHLPP2 or FOXO1 expression in bladder cancer tissues. C. The expression level of PHLPP2 or FOXO1 protein in bladder cancer cells transfected with miR-135a or miR-135a inhibitor, respectively, compared with control cells (shown as NC), by Western blotting analysis. α-Tubulin was used as a loading control. D. PHLPP2 or FOXO1 luciferase reporter activity in the indicated cells, measured by luciferase assay. pRL-TK Renilla luciferase was used as the normalization control; Bars represents the mean ± SD of three independent experiments. *P <0.05.