ROS-mediated PP2A activation increases phosphorylation of PTEN.
(A) PP2A activity was measured in DU145 and MEFs (*P < 0.05, analyzed by Mann-Whitney test), and PP2A-C IP was analyzed by Western blot. (B–D) Western blot analysis of indicated DU145 cells treated with (B) 1 μM okadaic acid for 2 hours, (C) 5 nmol PP2A-A siRNA for 42 hours, and (D) 5 nmol PP2A-C siRNA for 42 hours. (E) PP2A-A IP from DU145 and MEFs was analyzed by Western blot. (F) PP2A activity and Western blot of GSK3B IP in DU145 cells treated with 1 μM okadaic acid and 10 μM FTY720 for 2 hours (*P < 0.05, n = 3, analyzed by Mann-Whitney test). All Western blots were quantified using ImageJ, and the values represent relative immunoreactivity of each protein normalized to respective loading control. Data are presented as mean ± SEM (A and F).