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. 2020 Apr 24;11:2027. doi: 10.1038/s41467-020-15656-4

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© The Author(s) 2020

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. 8 — a Fate-mapping study showing that mGfap-Cre-mediated EYFP was expressed in GFAP⁺ astrocytes but not in Sox10⁺ oligodendroglial lineage cells or NeuN⁺ neurons in the spinal cord at P60. WM, white matter, GM, gray matter, Ctx, cortex. EYFP was identified as S100β⁺ astrocytes in the cerebral Ctx. Scale bars = 20 μm. b Representative images of Laminin immunostaining in mGfap-Cre, Vhl^fl/fl mutants and non-Cre control mice at P30. Scale bars = 100 μm. c Percentage of Laminin-occupying area among total area at P30. Two-tailed Student’s t test, Welch’s corrected t₍₄₎ = 11.92 spinal crd, t₍₇₎ = 10.12 cerebral cortex. n = 4 Ctrl, 5 VHL cKO. d RT-qPCR assay of endothelial Pecam1 at P30. Two-tailed Student’s t test, Welch’s corrected t_(4.046) = 8.564 spinal cord, Welch’s corrected t_(4.490) = 6.706 forebrain. n = 5 each group. e immunostaining showing that endogenous mouse IgG is restricted to Laminin⁺ blood vessels (arrowheads) in the early adult spinal cord of mGfap-Cre,Vhl^fl/fl mutant and control mice at P47. Scale bars = 10 μm. f RT-qPCR assay of the mRNA levels of HIFα target gene Glut1, Hk2 and Ldha in P30 spinal cord. Two-tailed Student’s t test with Welch’s correction, t_(4.099) = 7.947 Glut1, t_(4.243) = 9.636 Hk2, t_(4.085) = 9.025 Ldha. n = 4 Ctrl, 5 VHL cKO. g RT-qPCR assay of the mRNA levels of Wnt/β-catenin target genes Axin2 and Notum at P30. Two-tailed Student’s t test, Welch’s corrected t_(4.760) = 7.296 spinal cord Axin2, t₍₈₎ = 2.902 forebrain Axin2, Welch’s corrected t_(4.149) = 4.850 spinal cord Notum, t₍₈₎ = 4.994 forebrain Notum. n = 5 each group. h Immunostaining of active β-catenin in the spinal cord of mGfap-Cre, Vhl^fl/fl mutants and non-Cre control mice at P30. Scale bars = 10 μm. i Double immunostaining of active β-catenin and PECAM1 in the spinal cord of mGfap-Cre, Vhl^fl/fl mutants at P30. Arrowheads point to double-positive cells. Blue is DAPI nuclear staining. Scale bars = 10 μm. Data are shown as mean ± s.d. Source data of c, d, f, g are provided as a Source Data file.