Effect of iNOS on Bcl-2 S-nitrosylaiton, ubiquitination and protein expression and caspase signal pathway.

<p>(A) Time course analysis of iNOS expression in Hela, A375 and 7860 cells treated with ZD55-IL-24 (20 MOI). (B) Hela, A375 and 7860 cells were transfected with iNOS-siRNA (100 nM) for 12 h, then treated with ZD55-IL-24 for 12 h. Effect of iNOS-siRNA on iNOS expression was analyzed by Western blotting. (C) Effect of iNOS-siRNA on Bcl-2 protein expression was analyzed by Western blotting. (D) Effect of iNOS-siRNA on Bcl-2 S-nitrosylation were detected by immunoprecipitation using anti-Bcl-2 antibody and then followed by immunoblotted with anti-S-nitrosocysteine antibody. (E) Effect of iNOS-siRNA on ubiquitin-Bcl-2 was detected by immunoprecipitation using anti-Bcl-2 antibody and then followed by immunoblotting with anti-ubiquitin antibody. (F) Effect of iNOS-siRNA on caspase-9, caspase-3 and PARP in Hela cells was detected by Western blotting. The corresponding bands were scanned and the optical density (O.D.) was determined as the fold change versus control group. Data are means±standard deviation (S.D.) from three independent experiments (n = 3). *<i>p</i><0.05 versus control group; ^#<i>p</i><0.05 versus scrambled siRNA group.