Fig.1. EC-inducible Ccm3 deletion (Ccm3^ECKO) mice develop CCM lesions.

a–c. CCM lesion quantification in WT (Pdcd10^fl/fl) and Ccm3^ECKO (Cdh5CerERT2;Pdcd10^fl/fl). (a) H&E staining of cerebellum sections. Representative images of small lesions (arrows) and large lesions (asterisks) are shown. Number of total lesions (b) and lesions with different sizes (c) in Ccm3^ECKO mice were quantified (n=10). ns: non-significant; **P<0.01; ***P<0.001 (two-way ANOVA). d–g. Vascular leakage in CCM lesions. (d) Images for fresh brain tissue of WT and Ccm3^ECKO mice at P10. (e–f) P10 pups were perfused with FITC-dextran (2000 kDa) and brain sections were subjected to immunostaining with anti-CD31. Vascular leakage is indicated by arrows in panel e. CCM lesions are indicated by asterisks whereas arrowheads indicate a normal granule cell layer within each lobule of cerebella in WT mice (f). (g) Cerebellum permeability was measured by Evan’s blue dye (EBD) assay in WT and Ccm3^ECKO mice (n=10). **P<0.01 (unpaired two-tailed Student’s t-test). h–j. Lesions in retinas. P10 retinas from WT and Ccm3^ECKO pups were stained with EC markers CD31 (green) and isolectin B4 (red). CCM lesions with disorganized vasculatures are indicated by asterisks in confocal images of h (20×) and i (80×). Lesion areas (% of total retina) are quantified by Image J (j). n=10, **P<0.01 (unpaired two-tailed Student’s t-test). Error bars indicate s.e.m. Scale bars: a: 400 μm; f, h: 100 μm; d–e: 2 mm; i: 25 μm.