Figure 4
Effect of DPI, Cyt c, anti-NOX4 antibody, and AgNO 3 on insulin- and NSAID-mediated H 2 O 2 generation through a NOX4 system in isolated plasma membranes from rat adipocytes. Freshly prepared membranes were incubated for 10 min at 37°C in the activation buffer enriched with 4% BSA and 10−5 M GTPγS. Activation buffer was additionally supplemented with the following: none (control), or 5 ×10−5 M DPI, or 8 × 10−5 M Cyt c, or 1:6,400 dilution anti-NOX4 antibody, or 3 × 10−5 M AgNO3. In each set of experiments, 10−8 M insulin or 10−6 M of each NSAID was added to the incubation mixture. The H2O2 assay was conducted as described in Experimental Procedures. Each bar represents the average ± Standard error (SE) of three to six independent experiments. The letters (a, b, c, d, e, f) represent a significant difference among groups with different treatments by employing one-way Analysis of variance (ANOVA) at p <0.01. Groups with the same letter did not show a significant difference at p <0.05.