Figure 6. Actein potentiates ROS/JNK, and suppresses AKT pathway.

(A) BIU-87 and T24 cells were treated with different doses of ACT for 24 h, and then all cells were collected for DCF analysis to calculate the generation of ROS using fluorescent microscope. (B) BIU-87 and T24 cells were stained with MitoSOX Red after various treatments, and then the ROS levels were examined through confocal microscope. (C, D) The bladder cancer cells were pre-treated with ROS scavenger of NAC (5 mM) for 2 h, and then all cells were subjected with or without ACT (10 uM) for another 24 h, followed by DCF and MitoSOX Red staining using confocal microscope. (E) BIU-87 and T24 cells were treated with ACT for 24 h, and then p-AKT, p-mTOR, p-p38, p-JNK, p-STAT3 and p-JAK2 protein levels were evaluated using western blot analysis. Data are represented as mean ± S.E.M.