Effect of GSK-3β inhibition on serum-starvation-induced apoptosis.

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Effect of GSK-3β inhibition on serum-starvation-induced apoptosis. Effect of GSK-3β inhibition on serum-starvation-induced apoptosis. (A) C2C12 cells were co-transfected with M-cadherin-targeted siRNA and one of the plasmids (myrAkt, GSK-3βK85R or empty vector) for 48 hours, then serum-starved for 48 hours. DNA fragmentation was measured by ELISA. †P<0.05 vs M-/EV. (B) C2C12 cells were transiently transfected with M-cadherin RNAi (M-), or a non-targeted scrambled siRNA (SiCON). These were compared with normal non-transfected C2C12 cells (NC). The cells were treated with 20 μM TDZD-8 or DMSO for the last 12 hours of siRNA transfection. 48 hours after transfection, the treated and control cells were serum starved for 48 hours. The cells were harvested and DNA fragmentation was assessed by cell death ELISA. Each experiment was repeated three times. †P<0.05 vs SiCON/TDZD8 and M-/DMSO. (C) Mitochondria were isolated from each experimental group and stained with JC-1. The ratio of JC-1 orange to green staining was analyzed using a FACSCalibur system to measure the mitochondrial membrane potential. †P<0.05 vs M-/DMSO. All experiments were repeated three times. *P<0.05 vs NC or SiCON. Yan Wang et al. J Cell Sci 2011;124:3835-3847 © 2011.