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Published in final edited form as: Free Radic Biol Med. 2012 Jul 13;53(5):1181–1191. doi: 10.1016/j.freeradbiomed.2012.07.002

Fig. 9 — Knockdown of PRODH and Akt signaling pathway in prostate cancer cells. (A) RWPE-1 cells were treated with and without proline (5 mM) for 12 h and then incubated with 0.5 mM H₂O₂ for 3 h in serum free medium. Percent cell survival was estimated using the MTS cell viability assay. (B) Survival rates of PC3 cells transfected (48 h) with control siRNA (siCon) and PRODH siRNA (siPRODH) and incubated with different concentrations of H₂O₂ (0–10 mM) for 3 h. (C) Western blot analysis of PRODH in PC3 cells transfected with siCon (50 nM) and siPRODH (50 nM) for 48 h. VDAC is shown as a control. (D) Western blot analysis of P-Akt (T308 and S473), P-FoxO3a (T32), and β-actin in PC3 cells transfected with siCon and siPRODH (48 h). (E) Quantification of P-Akt-T308 and P-Akt-S473 relative to total AKT (P-Akt/Akt) and P- FoxO3a-T32 relative to β-actin. Each value represents mean ± SD of separate experiments (n = 4) (* P < 0.05).

Fig. 9 — Knockdown of PRODH and Akt signaling pathway in prostate cancer cells. (A) RWPE-1 cells were treated with and without proline (5 mM) for 12 h and then incubated with 0.5 mM H₂O₂ for 3 h in serum free medium. Percent cell survival was estimated using the MTS cell viability assay. (B) Survival rates of PC3 cells transfected (48 h) with control siRNA (siCon) and PRODH siRNA (siPRODH) and incubated with different concentrations of H₂O₂ (0–10 mM) for 3 h. (C) Western blot analysis of PRODH in PC3 cells transfected with siCon (50 nM) and siPRODH (50 nM) for 48 h. VDAC is shown as a control. (D) Western blot analysis of P-Akt (T308 and S473), P-FoxO3a (T32), and β-actin in PC3 cells transfected with siCon and siPRODH (48 h). (E) Quantification of P-Akt-T308 and P-Akt-S473 relative to total AKT (P-Akt/Akt) and P- FoxO3a-T32 relative to β-actin. Each value represents mean ± SD of separate experiments (n = 4) (* P < 0.05).