Figure 6. Impaired hemostasis, defective arterial thrombus formation, and protection from ischemic brain infarction in Bin2^fl/fl,Pf4Cre mice.

(A) Adhesion and thrombus formation of platelets on collagen was assessed in a flow adhesion assay at a wall shear rate of 1700/s. Representative images and the quantification of the surface coverage and the relative thrombus volume are shown (n = 4). Representative example of 3 independent experiments. Values are depicted as mean ± SD, and P values were calculated using the Mann-Whitney U test. *P < 0.05. Scale bar: 50 μm. (B) Tail bleeding times of WT and Bin2^fl/fl,Pf4Cre mice with or without acetylsalicylic acid (ASA) treatment (1 mg/kg i.v.) 15 minutes before the start of the experiment. Each symbol represents 1 animal. Kruskal-Wallis test (P value: 0.0022) followed by Dunn’s multiple-comparisons post hoc test; *P < 0.05, **P < 0.01. (C and D) Time to stable vessel occlusion of WT and Bin2^fl/fl,Pf4Cre mice without (C) or with (D) ASA treatment (1 mg/kg i.v.) 15 minutes before the start of the experiment. The abdominal aorta was injured by firm compression with forceps, and blood flow was monitored for 30 minutes. Mann-Whitney U test was performed to compare mean occlusion times of occluded vessels. To compare frequency of occluded and nonoccluded vessels, Fisher’s exact test was used. *P < 0.05, ***P < 0.001. (E–I) WT and Bin2^{fl/fl Pf4-Cre} mice were subjected to 60 minutes of transient middle cerebral artery occlusion (tMCAO) n = 10. (E) Representative images of 3 coronal brain sections stained with TTC. (F) Infarct volumes, (G) number of occluded ipsilateral vessels (2 slides per animal analyzed), (H) number of neutrophils, and (I) CD11b⁺ cells per section (2 slides per animal analyzed) were investigated after 24 hours. Values are depicted as mean ± SD, and P values were calculated using the Mann-Whitney U test. **P < 0.01, ***P < 0.001.