Fig. 2
Forced expression of miR-424 attenuates CDDP-mediated apoptotic cell death of bladder cancer cells. a Forced expression of miR-424. T24 and 5637 cells were transfected with the empty vector or with the expression vector for miR-424. Forty-eight hours after transfection, the expression level of miR-424 was examined by qRT-PCR. b miR-424 prohibits CDDP-induced proteolytic cleavages of PARP and caspase-3. Five thousand six hundred thirty-seven (left panels) and T24 (right panels) cells were transfected and treated as in (a). Twenty-four hours after treatment, cell lysates were analyzed by Western blotting with the indicated antibodies. β-actin was used as a loading control. c Overexpression of miRNA-424 mimics. Five thousand six hundred thirty-seven and T24 cells were transfected with the negative control (NC) or with miR-424 mimics. Forty-eight hours after transfection, the expression level of miR-424 was examined by qRT-PCR. d miR-424 mimics prohibit CDDP-induced apoptotic cell death. T24 (left panel) and 5637 (right panel) cells were transfected with the scrambled oligo or with miRNA-424 mimics followed by CDDP exposure (10 μM). Twenty-four hours after treatment, cells were subjected to FACS analysis. e miR-424 inhibitor reduces the expression level of miR-424. Five thousand six hundred thirty-seven and T24 cells were transfected with the control or with miR-424 inhibitor. Twenty-four hours after transfection, the expression level of miR-424 was examined by qRT-PCR. f miR-424 inhibitor augments CDDP-induced proteolytic cleavages of PARP and caspase-3. T24 cells were transfected with the scrambled oligo or with miRNA-424 inhibitor, and then exposed to CDDP (10 μM). Twenty-four hours after treatment, cell lysates were prepared and analyzed by Western blotting with the indicated antibodies. β-actin was used as a loading control. g CDDP-mediated apoptotic cell death is further enhanced by miR-424 inhibitor treatment. T24 (left panel) and 5637 (right panel) cells were transfected and treated as in (e). Twenty-four hours after treatment, cells were processed for FACS analysis. The data are presented as the mean ± S.D. of triplicate experiments. *P < 0.05, **P < 0.01