Fig. 3

WZ35 induced cell apoptosis through ER stress-mediated CHOP expression in PC-3 cells. a–b The time-course expression of ER stress markers induced by WZ35. Cells were treated with WZ35 (10 μM) at different time interval as indicated, then cells were processed for western blot analysis or RT-qPCR for expression of ER stress markers such as p-PERK, p-eIF2α, ATF4, and CHOP as described in methods, and GAPDH or β-actin was used as a loading control. c The mRNA expression of CHOP in cells treated with WZ35. Cells were treated with WZ35 at different concentrations as indicated for 12 h, then the mRNA expression of CHOP was detected with RT-qPCR assay as described in methods, and β-actin was used as a loading control. d–g The effects of CHOP-siRNA on cell apoptosis induced by WZ35. Cells were transfected with Ctrl siRNA or CHOP siRNA for 24 h as described in methods. Then the transfected cells were treated with or without WZ35 (10 μM) for 24 h. Then the cells were processed for western blot analysis for CHOP expression (d) or processed for CHOP immunofluoresence staining (e) or stained with Annexin V-FITC/PI followed by flow cytometry analysis (f) or processed to western blot analysis for cleaved-caspase3 expression (g), as described in methods. The statistic data were presented as mean ± S.E from three independent experiments. *, p <0.05, versus Ctrl siRNA + WZ35 group