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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. 4 — a Scheme of study design. The lungs of tamoxifen-inducible EC-Atm^-/-, EC-Bmpr2^-/- and control mice (n = 5) after reoxygenation were harvested, enzymatically digested to obtain single-cell suspensions, and pulmonary endothelial cells (CD45⁻ CD31⁺ cells) were collected using magnetic beads. Total RNA of pulmonary EC from n = 5 mice was used for bulk RNA sequencing. Schema created with BioRender.com. b Venn diagram showing the number of genes downregulated following reoxygenation in lung EC of EC-Atm^-/- or EC-Bmpr2^-/- mice, compared to control mice [adjusted P-value (Padj) <0.05]. Gene Ontology (GO) analysis was performed for 547 commonly downregulated genes to reveal the top GO pathways. P-values were adjusted using the Benjamini–Hochberg method for multiple comparisons by one-sided Fisher’s exact test. c, d Volcano plots displaying DEGs (differentially expressed genes) following reoxygenation in EC-Atm^-/- or EC-Bmpr2^-/- compared to control mice (c, d respectively). Angiogenesis genes (Foxf1, Vegfr2, Acvrl1, S1pr1, Eng, Bmpr2, Id1, Klf4, Cdh5, Cldn5, Ednrb), Trp53inp2, a gene downstream of p53, and Foxf1 targeting as well as DNA repair genes (Fancc, Smarca2, Rad21, Rrm2b, Cep164) are indicated on the plots. 2-sided Wald test was performed. P-values were adjusted to get Padj using the Benjamini-Hochberg method. e Foxf1 expression in lung EC of EC-Atm^-/-, EC-Bmpr2^-/- and control mice following reoxygenation. n = 3 biological replicates. f Immunoblot and densitometry of FOXF1, CLDN5, VEGFR2, P53 and ATM in pulmonary EC of EC-Atm^-/-, EC-Bmpr2^-/- and control mice after reoxygenation. n = 3 biological replicates. e, f Bars represent mean ± S.E.M. P values determined by one-way ANOVA with Holm-Sidak posthoc test. ns, not significant. Source data are provided as a Source Data file.