Fig.5. UNC13-deficiency rescues CCM phenotypes in Ccm3^ECKO mice.

WT (Pdcd10^fl/fl), Unc13b^−/−, Ccm3^ECKO (Cdh5CerERT2;Pdcd10^fl/fl) and DKO (Ccm3^ECKO;Unc13b^−/−) pups were fed with tamoxifen from P1 to P3 to induce deletion of CCM3. Cerebella were harvested at P10, and frozen sections were stained as indicated. a. Images for fresh brain tissue. Arrows indicate lesions. b–c. H&E staining and lesion quantifications. d–e. Co-staining for NG2 with CD31 and quantification of NG2⁺ coverage on microvessels. f–g. Co-staining for ANGPT2 with CD31 and p-TIE2 with CD31 and quantification of p-TIE2-positive ECs. Arrowheads indicate normal vessels and asterisks indicate lesions. h–i. Brain tissues were collected and ANGPT2 levels were determined by ELISA (h) and by Western blotting (i). j. Mouse brain microvascular ECs were isolated and ANGPT2 secretion from culture supernatants was measured by ELISA. n=10, *P <0.05; **P <0.01 (one-way ANOVA). Error bars indicate s.e.m. Scale bars: a: 2 mm; b (top): 400 μm; b (bottom), d, f: 100 μm.