Vps34 plays an important role in thrombosis via its kinase activity. (A) Tail bleeding time (n = 30 mice of each genotype) was measured as described in “Methods.” (B) The thrombotic response of mice to carotid injury after exposure to 7.5% ferric chloride for 3 minutes was assessed by a flow probe. Graph represents the percentage of mice without occlusion 30 minutes after injury (n = 15 mice of each genotype; P = .0002 according to 1-sample Student t test). (C-D) DiOC6-labeled platelets in mouse whole blood were perfused through collagen-coated microcapillaries at a physiological arterial shear rate of 500 s−1 (C) or at an arteriolar shear rate of 1500 s−1 (D). Scale bar represents 20 µm. The surface covered by fluorescent platelets and thrombus volume were analyzed using ImageJ software (mean ± SEM; n = 8 mice of each genotype; *P < .05; **P < .01 vs WT according to 2-tailed Student t test and 1-sample Student t test). (E) Healthy human donor whole blood was treated with 1 µM VPS34-IN1, 1 µM SAR405, or vehicle (dimethyl sulfoxide) for 1 hour. Then, DiOC6-labeled whole blood was perfused through collagen-coated microcapillaries at a physiological shear rate of 1500 s−1. Scale bar represents 20 µm. The surface covered by fluorescent platelets and thrombus volume were analyzed using ImageJ software (mean ± SEM; n = 3-5 healthy donors depending on the inhibitor; *P < .05; **P < .01 vs vehicle according to 2-way ANOVA and 1-sample Student t test).
