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. 2021 Mar 10;12:1565. doi: 10.1038/s41467-021-21865-2

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© The Author(s) 2021

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 A total of five clusters of endothelial cells were identified in developing lungs. Cell populations are colored as indicated by the legend. b UMAP plots of principal identifiers of different types of endothelial cells. The intensity of expression is indicated by purple coloring. c Heatmap of top 5 most differentially expressed genes across endothelial clusters. The intensity of expression is indicated as specified by the color legend. d UMAP plots depicting cell identity in regards to developmental time points in normally (purple) and aberrantly (green) developing lung endothelium. The intensity of expression is indicated as specified by the color legend. Violin plots depicting changes in gene expression of Pecam and Inhba in normoxia (21% O₂, green) and hyperoxia-specific (85% O₂, red) at P14. e Representative images from fluorescent RNA in situ hybridization for hyperoxia-specific marker Inhba (pink) and pan-endothelial marker Pecam (green) in developing mouse lungs. Magnification: 40×. Scale bar = 40 µm. Two 14-days old animals/group were analyzed. f Representative images and quantitative analysis of fluorescent RNA in situ hybridization for hyperoxia-specific marker INHBA (red) and pan-endothelial marker PECAM (white) in BPD patients’ and donors’ lungs. Samples from five BPD patients and two donor lungs were analyzed. For quantitative analysis, cells in 15 randomly chosen fields of view/sample were analyzed. Magnification: 40×. Scale bar = 40 µm. g Dot plot depicting hyperoxia-induced changes in the expression of inflammatory, anti-angiogenic and protective genes in capillary cells at P14. The intensity of expression is indicated by the color legend. Size of the cell population expressing the gene of interest is indicated by the size of the circle as specified by the legend. h Hyperoxia-impacted signaling pathways in gCap (pink), aCap (yellow), and Vein (blue) clusters, as identified by gene set enrichment analysis (GSEA). All terms are significantly enriched (adjusted p value < 0.05) and normalized enrichment scores (NES) are shown. NES values were computed by gene set enrichment analysis on fold change-ranked genes. Expression values in Heatmap represent Z-score-transformed log(TP10k + 1) values. Expression levels in UMAP plots and Dotplots are presented as log(TP10k + 1) values. Log(TP10k + 1) corresponds to log-transformed UMIs per 10k.