Increased rate of hemorrhage and mortality after tMCAO in DKO mice. (A) Mortality rates 24 hours after tMCAO of WT and DKO mice, as well as DKO mice transfused with WT platelets. (B) Representative images show microbleedings in TTC-stained brain sections (left) or intraparenchymal (middle) and subarachnoid (right) bleedings (arrows) in native brain sections of nonsurviving double-deficient mice after stroke. (C) Representative images of coronal sections stained with TTC 24 hours after tMCAO of WT and surviving DKO mice. Infarcted areas are shown in white. (D) Planimetric analysis was used to quantify the infarct volume. Results are presented as mean ± SD. n = 20 (WT), n = 10 (DKO), and n = 4 (DKO + WT platelets) mice. Bederson score (E) and grip test (F) were used to analyze neurologic outcome in the surviving animals 24 hours after tMCAO. Each symbol represents 1 mouse. n = 20 (WT), n = 10 (DKO), and n = 4 (DKO + WT platelets) mice. (G) Quantification of fluorescein isothiocyanate-dextran extravasation in brains from naïve WT and DKO animals. n = 5 mice per group. (H) Immune fluorescent staining of tight-junction proteins occludin and claudin-5 in brain sections from naïve WT and DKO animals using a 40× objective. Representative images from n = 3 mice per group. Scale bar 40 μm. ***P < .001. DAPI, 4,6 diamidino-2-phenylindole; MFI, mean fluorescence intensity; n.s., not significant; SD, standard deviation.