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. 2013 Jan;344(1):286–294. doi: 10.1124/jpet.112.199067

Fig. 6.

Fig. 6.

The effect of APAP on protein sulfhydryls in mouse liver and the restoration of reversibly oxidized protein sulfhydryls. Mice (n = 3–5) were administered a single oral gavage dose of 0.5% methylcellulose vehicle (Cntl), APAP low dose (150 mg/kg), or APAP high dose (300 mg/kg) and total liver protein (24 hours) was isolated and resolved on a 12% SDS-PAGE gel. Protein thiols were labeled with maleimide-IRDye before loading on the gel. (A) The high-dose samples had decreased protein thiols. (B) The same samples were reduced with TCEP, desalted, and then labeled with maleimide-IRDye before loading on the gel. (C) The same samples were resolved on a duplicate gel followed by Coomassie Blue staining to confirm equal loading of protein between samples. The levels of protein sulfhydryl loss were quantified with a near-infrared scanner. (D) These results show that APAP induces protein sulfhydryl oxidation and the majority of the oxidation is reversible with a thiol-reducing agent (TCEP). *Protein thiols of 300 mg/kg group were statistically significantly decreased (P < 0.05) compared with the control and TCEP-treated samples.