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. Author manuscript; available in PMC: 2011 Nov 30.
Published in final edited form as: Nitric Oxide. 2008 Apr 24;19(2):146–151. doi: 10.1016/j.niox.2008.04.019

Figure 4. Effects of NO on S-nitrosylation and ubiquitination of Bcl-2.

Figure 4

A, Effect of NO modulators on Bcl-2 S-nitrosylation in Cr(VI)-treated cells. Human lung epithelial H460 cells overexpressing myc-Bcl-2 were either left untreated or pretreated with SNP (500 μg/ml), DPTA NONOate (400 μM), AG (300 μM), PTIO (300 μM), or DTT (10 mM) for 1 h. The cells were then treated with Cr(VI) (20 μM) for 3 h and cell lysates were prepared for immunoprecipitation using anti-myc antibody. The resulting immune complexes were analyzed by Western blot using S-nitrosocysteine antibody. Densitometry was performed to determine the relative S-nitrosocysteine levels after reprobing the membranes with anti-Bcl-2 antibody. B, Effect of NO modulators on Cr(VI)-induced Bcl-2 ubiquitination. Cells overexpressing myc-Bcl-2 were treated with the indicated test agents in the presence of lactacystin (10 μM) to prevent proteasome-mediated Bcl-2 degradation. Cell lysates were then immunoprecipitated with anti-myc antibody and the immune complexes were analyzed for ubiquitin. Data are mean ± S.D. (n = 4). *p < 0.05 versus Cr(VI)-treated controls. #p < 0.05 versus NO-modulated controls. (Reproduced from ref. 20 with permission from the Journal of Biological Chemistry).