Fig. 3.

Knockdown of MSH2 increases cisplatin resistance in bladder cancer cell lines. Two shRNA constructs targeting MSH2 and one nontargeting shRNA control were transduced into MGHU4 and 253J bladder cancer cell lines. (A) Western blot showing the level of knockdown of MSH2. (B) MGHU4 and (C) 253J cell lines were treated with the indicated doses of cisplatin for 48 h, and cell viability was measured using an ATP-based assay (mean ± SEM of three independent experiments). The GI50 (μM) of cell line was calculated with using a four-parameter nonlinear regression fit using least squares. (D) MGHU4 and (F) 253J cells were treated with 10 and 15 μM cisplatin, respectively, or vehicle. Cumulative caspase activation (green cells) was measured and normalized to cell confluency. Representative images of (E) MGHU4 and (G) 253J cells are shown at the 24 h time point. Statistical significance of Figures 3D and 3F were determined using a repeated-measure one-way ANOVA (independent variable is knockdown). Data represents the mean ± SEM of a representative experiment (of three experiments). ANOVA = analysis of variance; SEM = standard error of the mean. **** p < 0.0001.