Fig. 6.

Collagen-dependent thrombus formation at arterial shear requires both Nox1 and Nox2. Hirudin-anticoagulated whole blood from wildtype (Wt) or Nox2 knockout (KO) mice was diluted 1:2 with modified HEPES-Tyrode's buffer, then pre-incubated with vehicle control (0.5% DMSO), Nox1 inhibitor (5 µM ML171) or aspirin (500 µM). Samples were perfused over a collagen-coated microfluidic channel at a constant shear of 1500 s⁻¹ for 6 min, followed by washing in modified HEPES-Tyrode's buffer for 5 min with subsequent fixation in 3.7% formaldehyde. (A) Representative fluorescent, end point images of platelet thrombi from Wt (upper panel) and Nox2 KO (lower panel) mice following vehicle control/inhibitor treatment. (B) Platelet surface coverage (left panel) and thrombus volume (right panel) are expressed as mean±SEM, n=5 (n=3 for aspirin), ^⁎P<0.05, ^⁎⁎P<0.01 by two-way ANOVA with Bonferroni analysis.