Fig. 3

ASC-J9® increases ROS and decreases GCLC levels in PCa cells. a Western Blots show higher ATF3 expression with ASC-J9® treatment (left panels) than with Enz treatment (right panels) in C4–2 (upper) and CWR22Rv1 (lower) cells. b Western Blots show 5 mM NAC treatment can suppress ASC-J9® induced ATF3 expression in C4–2 (upper) and CWR22Rv1 (lower) cells. c Western Blot shows ASC-J9® induced ATF3 expression in AR negative PCa PC-3 cells. d DHE staining assay shows ROS level increase in PC3 after ASC-J9® treatment and 5 mM NAC treatment could suppress DHE signal. e GSH assay to reveal ASC-J9® can decrease GSH concentration in C4–2 and CWR22Rv1 cells. f The qRT-PCR of GSH upstream genes GCLC was decreased significantly in C4–2 and CWR22Rv1 cells when treated with ASC-J9®. For a, c-d, quantification is at the right. Data represent the mean ± SD except quantitative real time-PCR represent the mean ± SEM. *p < 0.05, **p < 0.01, ns = not significant, by unpaired Student t test