Changes in the levels of CYP2E1 protein levels, CYP2E1 activity, MDA+HAE concentration, hepatic protein oxidation, nitration and glycosylation parameters(A–C) Equal amounts of whole liver lysates from different groups were used to determine: (A) CYP2E1 protein levels (upper panel) and p38 as a loading control (lower panel) by immunoblot analysis; (B) CYP2E1 activity evaluated by measuring the rate of PNP oxidation to p-nitrocatechol, and (C) hepatic MDA+HAE as a marker for lipid peroxidation. *Significantly different from all other groups. N.D.: not detected. (D) Protein oxidation was determined using Oxyblot analysis of total cells lysates from all four groups (upper panel) or negative control (lower panel). (E) Equal amounts of whole liver lysates (40 μg/well) from different groups were used to determine protein nitration using anti-3-NT antibody (upper panel) or p38 as a loading control (lower panel). (F) Total cell lysates were used to evaluate the levels of AGE (top), RAGE (middle), or p38 as a loading control (bottom) from all four groups using immunoblot analysis. (G) Whole liver homogenates from different groups were used to determine the levels of glycoproteins (arrows indicate increased levels of glycoproteins). (H) Equal protein loading for the samples analyzed for glycoproteins was verified by staining with silver.
