Fig. 6

a Double immunofluorescence labeling of laminin (blue) and SMA (red) is used to illustrate the extravasation of FITC-albumin (green) at the level of arteries, capillaries and veins. Images are obtained from the ischemia-affected striatum of 2 h pMCAO and 4 h pMCAO mice. Scale bar: 10 μm. b Further, the extent of FITC-albumin related BBB breakdown was analyzed along different segments of the vascular tree in 2 h and 4 h pMCAO mice. Here, the mean fluorescence intensity of FITC-albumin extravasations, the area of the extravasations per type of vessel and the relative contribution per field of view (FOV) were analyzed (n = 4, ANOVA followed by Bonferroni’s multiple comparison test). c Analysis of the mean score of ultrastructural damage at the level of electron microscopy in ischemia-affected arterial vessels and adjacent capillaries. In line with the results shown in Fig. 4, the mean scores of ultrastructural damage are significantly increased from 30 min to 4 h of MCAO. Importantly, arterial and capillary endothelial cells exhibit comparable scores of ultrastructural damage in the ischemia-affected striatum and the cortex. Compared to arterial and capillary endothelial cells, arterial smooth muscle cells of the vascular wall are significantly less affected after 2 and 4 h of ischemia, predominantly showing a cellular edema (‘score 1’), only. Comparison between 4 h pMCAO and tMCAO animals did not provide statistically significant differences. 30 min, 1 h, 2 h pMCAO and 4 h tMCAO: n = 4; 4 h pMCAO: n = 5; ANOVA followed by Bonferroni’s multiple comparison test. (b & c) Data are given as means. Error bars indicate SD