Platelet aggregation and accumulation/thrombi formation ex vivo are similar in WT and Nox2-deficient male or female mice. Washed platelets from male and female WT (+/y or +/+) or Nox2-deficient (−/y or −/−) mice were activated with 25 ng/mL convulxin (A-C) or 0.02 U/mL thrombin (D-F). Representative aggregation tracings for convulxin (A) and thrombin (D). Quantitative data for the percentage light transmission at different time points for convulxin in male (B) and female (C) mice and for thrombin in male (E) and female (F) mice. Washed platelets from male and female WT and Nox2-deficient mice were perfused over a collagen surface for 5 minutes in a microfluidic flow chamber at a shear rate of 2000/s. (G) Representative images of platelet accumulation after 5 minutes of perfusion for male and female mice. (H) The time course of accumulation of platelets/thrombi development was calculated as the surface area covered by platelets in a fixed field. (I) Total thrombi area after 5 minutes of perfusion was calculated as the average surface area covered by platelets in 5 representative fields. Data are presented as mean ± standard error (n = 5-7 mice in each group). Platelet aggregation data were analyzed using multiple Student t tests with Holm-Sidak test for comparison between the groups. Platelet accumulation over time was compared using 2-way ANOVA repeated measures, and total thrombi areas on collagen surface were analyzed by 1-way ANOVA, followed by the Tukey test for multiple comparisons. *P < .05 vs. +/y, ^$P< .01 vs. −/y.