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. Author manuscript; available in PMC: 2013 Jun 15.
Published in final edited form as: Biochem Pharmacol. 2012 Mar 24;83(12):1643–1654. doi: 10.1016/j.bcp.2012.03.010

FIGURE 8.

FIGURE 8

Superoxide induces ERK1/2 inactivation and cysteine sulfenic acid formation. A, HaCaT cells were treated with DOX (1.5 μM) for 6 h in the presence or absence of MnTBAP (50 μM), catalase (CAT, 7,500 units/ml), or DMTU (5 mM), and phospho-Bcl-2 (p-Bcl-2) and phospho-ERK1/2 (p-ERK1/2) expression was determined by Western blotting. Blots were reprobed with ERK1/2 antibody to confirm equal loading of the samples. B, cells were treated with various concentration of DOX (0-3 μM) for 2 h and cell lysates were prepared and immunoprecipitated with anti-ERK1/2 (lower) antibody. The immune complexes were analyzed for cysteine sulfenic acid (Cys-SOH) by Western blotting. C, cells were treated with DOX (1.5 μM) in the presence or absence of MnTBAP (50 μM), catalase (CAT, 7,500 units/ml), or DMTU (5 mM). Analysis of ERK1/2 Cys-SOH was performed as described. Plots are mean ± S.D. (n = 3). *, p < 0.05 versus non-treated control. #, p < 0.05 versus DOX-treated control.