Effect of TrxR1 on Bcl-2 S-nitrosylaiton, ubiquitination and protein expression and caspase signal pathway.

<p>(<b>A</b>) Time course analysis of TrxR1 expression in Hela, A375 and 7860 cells treated with ZD55-IL-24 (20 MOI). (<b>B</b>) Hela, A375 and 7860 cells were treated with ZD55-IL-24 for 24 h and then added with TrxR1 inhibitor auranofin (5 µM) for 6 h. Effect of TrxR1 inhibitor auranofin on TrxR1 protein expression was detected by Western blotting. (C) Effect of TrxR1 inhibitor auranofin on Bcl-2 protein expression in Hela, A375 and 7860 cells was detected by the western blot assay. (D) Effect of TrxR1 inhibitor auranofin on Bcl-2 S-nitrosylation was detected by immunoprecipitation using anti-Bcl-2 antibody and then followed by immunoblotting with anti-S-nitrosocysteine antibody. (E) Effect of TrxR1 inhibitor auranofin on Bcl-2 ubiquitination in Hela cells was detected by immunoprecipitation using anti-Bcl-2 antibody and then followed by immunoblotting with anti-ubiquitin antibody. (F) Effect of TrxR1 inhibitor auranofin on caspase-9, 3 and PARP was detected by Western blotting. β-actin was used as a loading control. 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 ZD55-EGFP; ^#<i>p</i><0.05 versus DMSO group.