Figure 2.
The amounts of superoxide-specific 2-OH-E+generated from the oxidation of DHE are similar in Nox2-deficient and WT mice. Bead-purified platelets (pooled) from male Nox2-deficient (−/y) or WT (+/y) mice were incubated with DHE (25 µM), followed by activation (no stirring) or aggregation (with stirring) with thrombin (0.05 U/mL) and convulxin (50 ng/mL). The superoxide-specific oxidation product 2-OH-E+ was quantitated in platelet pellets (intracellular signal) (A) and supernatant fractions (extracellular signal) (B) using HPLC coupled with electrochemical detection and authentic standards. Data are presented as mean ± standard error (n = 3 or 4 mice in each group). *P < .05 vs. RP. 2-OH-E+ generation within different experimental conditions (ie, RP, Act, or Agg) and genotypes was analyzed using 2-way ANOVA with the Tukey test. Act, activated platelets; Agg, aggregated platelets; RP, resting platelets.