Fig. 4
PRDX1 suppresses JNK activation in MFs (a) BALB/c MFs were analyzed for H2O2 scavenging using the luminol hypochlorite assay. (b) PRDX1 dimer formation and over-oxidation on Cys52 was analyzed in BALB/c MFs treating cells with H2O2 as indicated by immunoblotting under non-reducing conditions. (c and d) Decreased expression of PRDX1 in murine MFs results in PTEN oxidation and intra-disulfide formation in the presence of H2O2. BALB/c MFs were starved for 24 h in 0.25% FBS DMEM and stimulated with 100 μM H2O2 for 5 min in the presence of NEM and analyzed under non-reducing conditions by western blotting for PTEN. Scr: scramble, sh2: shPRDx1–2, and sh4: shPRDX1–4 (D) represents the average of relative oxidized to reduced PTEN levels in shPRDx1–2 and shPRDX1–4 expressing cells normalized to EV with STDEV (e) Prdx1−/− and Prdx1+/+ MFs were analyzed by immunoblotting for phosphorylation of c-jun, ATF2 and JNK (f) Spontaneously immortalized MFs isolated from 8-wk-old virgin female BALB/c mice were stimulated with 100 μM H2O2 for 5, 10 and 30 min before analyzed for JNK phosphorylation and JNK1 by immunoblotting