Figure 6.

AZD0530 treatment blocks the Src signaling pathway in the ischemic brains of EC-targeted miR-15a/16-1 cKO mice. EC-miR-15a/16-1 cKO mice and WT littermate controls were subjected to 1 h MCAO followed by 14d reperfusion. Vehicle (Veh) or AZD0530 (AZD, 20 mg/kg) was administered daily to both genotypes at 3-14d after MCAO by oral gavage. Total proteins were isolated from the ipsilateral cortex of ischemic brains, and western blotting was carried out to detect the expression of proteins in the Src signaling pathway. (a–g) representative western blotting images (a,e) and quantitative analysis (b,c,d,f,g) indicate endothelial-selective deletion of the miR-15a/16-1 cluster upregulated the Src signaling pathway by enhancing phosphorylated-Stat3 (Tyr-705) (b), phosphorylated-Akt (Ser-473) (c), phosphorylated-FAK (Tyr-397) (d), phosphorylated-p44/42 MAPK (Thr202/Tyr204) (f), and phosphorylated-p38 MAPK (Thr180/Tyr182) (g), whereas AZD0530 effectively downregulated the phosphorylation of these Src kinase downstream mediators (b,c,d,f,g) in the ischemic brains of EC-miR-15a/16-1 cKO mice 14d after reperfusion. Data are expressed as the mean ± SD. n = 4/group for (b,d,g). n = 5/group for (c). n = 6/group for (f). *p < 0.05, **p < 0.01, ***p < 0.001 as indicated. Statistical analysis was performed by one-way ANOVA followed by Bonferroni's multiple comparison tests.