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. 2019 Jun 27;10:2817. doi: 10.1038/s41467-019-10643-w

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 1 — Pdgfrb-CreERT2-mediated Rbpj deletion compromises brain vessels. a Pdgfrb-CreERT2 transgene and Cre-mediated recombination of Rbpj^lox allele. b Whole brains and coronal sections from control and Rbpj^iPC mice at P7 or P10 after tamoxifen administration (P1–P3). Scale bar, 2 mm. c Haemorrhagic index. p-values, Mann–Whitney U-test, n = 8. d Red blood cell (RBC) extravasation in brain parenchyma. HPF, hippocampal formation. p-values, one-way ANOVA and Tukey’s test, n = 5. e Vascular area and density in brain cortices. p-values, Student’s t-test, n = 6–7. f Confocal images of blood vessels (CD31⁺, white) in brain cortices. Arteries (A) and veins (V) are indicated. Scale bar, 100 μm. g Quantitation of vessel diameter in cortical blood vessels. p-values, Brown–Forsythe and Welch one-way ANOVA with Tamhane’s T2 test, n = 4. n.s., not statistically significant. h Arteriovenous malformation in brain superficial vasculature as revealed by the presence of latex in the caudal rhinal vein (arrowhead). Scale bar, 1 mm. i Confocal images of cortical blood vessels stained for GLUT1 (red, white) and PDGFRβ (white). Normal sprouts (dashed circles) replaced by blunt-ended capillaries (yellow arrowheads) in Rbpj^iPC mice. Scale bar, 50 μm. j Quantitation of sprouting in brain cortices. p-values, Welch’s t-test, n = 4. k Confocal images of blood vessels (ICAM2⁺, red) and EC nuclei (ERG⁺, white) in brain cortices. Scale bar, 100 μm. l Quantitation of EC proliferation in blood vessels of P7 mice. p-values, Welch’s t-test, n = 7–8. m Confocal images of cortex capillaries stained for ICAM2 (red), ERG (white), and GFP (green, recombined mural cells). Right panel shows single optical sections of boxed inset in Rbpj^iPC (left column). Recombined pericytes (white arrows) around tangled capillaries with superimposed EC nuclei (white arrowheads). Scale bar, 50 μm (left panels) and 10 μm (high magnifications). n RT-qPCR analysis in sorted ECs from brain cortices of P7 and P10 mice. p-values, Brown–Forsythe and Welch one-way ANOVA with Tamhane’s T2 test for Dll4 and Myc; one-way ANOVA with Sidak’s test for Esm1, n = 4–8. All images correspond to P10 mice unless otherwise indicated. Error bars represent s.e.m. Source data are provided as a Source Data file