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. 2023 Nov 21;14:7578. doi: 10.1038/s41467-023-43039-y

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© The Author(s) 2023

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. 5 — a Scheme of study design. The right lungs of tamoxifen inducible EC-Bmpr2^-/- (n = 3 mice) or control mice (n = 4 mice) after reoxygenation were harvested, pooled, and enzymatically digested to obtain single-cell suspensions. Pulmonary endothelial cells (CD45^- CD31⁺ cells) were collected using magnetic beads and analyzed by 10X single-cell RNA sequencing. Schema created with BioRender.com. b Uniform Manifold Approximation and Projection (UMAP) of tdTomato⁺ traced lung EC in EC-Bmpr2^-/- and control mice following reoxygenation (n = 27,828 cells). Pulmonary EC (CD45^-CD31⁺ cells) were selected using magnetic beads and single-cell RNA seq performed on tdTomato-positive cells (tdTomato gene expression > 0). EC clusters were identified using EC subpopulation markers: Ednrb for aerocytes; Bmx for artery EC; Aplnr for general capillary (gCap); Prox1 for Lymphatic EC; Slc6a2 for Vein EC. Numbers of cells in each subtype: Aerocytes, n = 3574 cells; Artery EC, n = 1798 cells; gCap, n = 17,920 cells; Lymphatic EC, n = 724 cells; Vein EC, n = 3812 cells. c Violin plots of the expression of the EC subpopulation markers Ednrb, Bmx, Aplnr, Prox1 and Slc6a2. a.u., arbitrary units. d UMAP in (a), separated to control (left; n = 15,359 cells) and EC-Bmpr2^-/- (right; n = 12,469 cells) mice. Numbers of cells in control mice: Aerocytes, n = 2232 cells; artery EC, n = 882 cells; gCap, n = 10,616 cells; Lymphatic EC, n = 324 cells; Vein EC, n = 1305 cells. Numbers of cells in EC-Bmpr2^-/- mice: Aerocytes, n = 1342 cells; artery EC, n = 916 cells; gCap, n = 7304 cells; Lymphatic EC, n = 400 cells; Vein EC, n = 2507 cells. e. UMAP of angiogenesis genes colored by expression levels in EC-Bmpr2^-/- or control mice following reoxygenation. Color scale: Purple, high expression; white, low expression. f Violin plots showing expression of angiogenesis genes in each pulmonary EC subpopulation of EC-Bmpr2^-/- or control mice following reoxygenation. a.u., arbitrary units. P values are determined by 2-sided Wilcoxon rank sum test. ns, not significant. Source data are provided as a Source Data file.