Oxidative Stress Induced by the Deubiquitinase Inhibitor b-AP15 Is Associated with Mitochondrial Impairment

Figure 5

Proteomic analysis of mitochondrial proteins. (a) Volcano plot showing log₂(fold change) versus values for proteins from isolated mitochondria prepared from HCT116 cells treated with DMSO or 1 μM b-AP15 for 6 h. (b) Top candidates with significant changes from (a) (, or ). (c) Volcano plot showing log₂(fold change) versus values for proteins from isolated mitochondria prepared from HCT116 cells treated with DMSO or 1 μM b-AP15 and 10 μM CpdA for 6 h. (d) Top candidates with significant changes from (c) (, or ). (e, f) Upper part: HCT116 cells were exposed to 0.5% DMSO, 1 μM b-AP15 in the presence or absence of 10 μM CpdA for 1, 3, and 6 h, as indicated. Extracts were prepared and subjected to immunoblotting using antibodies to Hsp60 and β-actin. (e, f) Lower part: mitochondria were purified from cells treated with 1 μM b-AP15 in the presence or absence of 10 μM CpdA and analyzed by shotgun proteomics. Data was normalized to control samples (treated with 0.5% DMSO).

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