Involvement of Cdh5 in miR-132 knockdown-induced defects of brain vascular integrity. (A-C) Effects of miR-132 knockdown on the expression of junction proteins. Representative blots (A) and summary (B) of Western blot analyses. The experiments were repeated 3-8 times. (C) Cdh5 immunostaining in 3-dpf Tg(Flk1:eGFP) larvae. (D and E) Effects of Cdh5 knockdown on intracranial hemorrhage (D) and DAPI leakage (E) in 3-dpf larvae. The experiments were repeated 6 times and at each time > 49 embryos were examined for each group in D. 23 control embryos and 24 morphants were analyzed in E. (F) Effect of miR-132 knockdown on vsg1 expression. The experiments were repeated three times. (G, H) Representative projected confocal images (G) and summary (H) showing no significant effect of miR-132 knockdown on the pericyte coverage of brain blood vessels. 3-dpf larvae generated by crossing Ki(pdgfrβ:Gal4) with Tg(UAS:GFP;Fli1:DsRed) were used, and GFP and DsRed are expressed in pericytes and ECs, respectively. 6 control embryos and 6 miR-132 morphants were analyzed. Scale bar, 25 μm (C), 50 μm (G). Error bars, SEM. n.s., not significant; *P < 0.05, **P < 0.01, ***P < 0.001 (unpaired two-tailed Student's t-test for B, D, E, F and H).
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