Fig.6. ANGPT2 neutralizing antibody blunts CCM lesion progression in Ccm3^ECKO mice.

WT, Ccm3^ECKO and DKO (Ccm3^ECKO;Unc13b^−/−) pups were fed with tamoxifen from P1 to P3 to induce CCM3 deletion. Pups were intraperitoneally injected with control IgG or ANGPT2 neutralizing antibody (10 μg/g body weight) from P2 and every other day thereafter. Cerebella were harvested at P10 in frozen sections. 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, VE-cadherin with CD31 and Claudin-5 with CD31, and quantifications of NG2, VE-cadherin and Claudin-5 coverage on microvessels. f. Co-staining for ANGPT2 with CD31. g. Brain tissues were collected and ANGPT2 levels were determined by ELISA. h–i. Co-staining for p-TIE2 with CD31 and quantification of p-TIE2-positive ECs. Arrowheads indicate normal or normalized vessels and asterisks indicate lesions. 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, h: 100 μm.