Fig. 4
T-96-mediated ROS accumulation induced ER stress. A DU145 and PC3 cells were exposed to the indicated concentrations of T-96 for 48 h, and then treated with DCFH-DA for another 30 min. Immunofluorescence analysis was performed to examine the green signal representing ROS generation. Green signal cells were counted and presented in the histogram. Scale bar: 100 μm. B T-96 induced accumulation of ubiquitinated proteins. DU145 and PC3 cells were treated with indicated concentrations of T-96 for 48 h, and the ubiquitination levels were then detected using western blot. C T-96 activated the UPR pathway in a dose dependent manner. UPR pathway-related proteins were detected using western blot. D T-96 enhanced intracellular Ca2+ concentration in a dose dependent manner. DU145 and PC3 cells were exposed to the indicated concentrations of T-96 for 24 h, and then cells were incubated with 5 µM Fluo-4/AM for 1 h and detected using fluorescence microscopy. Data are presented as the means ± SD. *P < 0.05 and **P < 0.01 compared with the control group