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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. 6 — a The relative proportion of myeloid (purple) and lymphoid (teal) cells in developing lungs was significantly impacted by hyperoxia exposure. n = 6 animals/group. Data are presented as means ± SD. Statistical analyses were performed with GraphPad Prism 8.0. Significance for each population at each time point was evaluated by unpaired, two-tailed Student’s t-test. P value = 0.0013 for Myeloid population, and 0.0086 for Lymphoid population. b A total of eight clusters of myeloid cells were identified in developing lungs. Cell populations are colored as indicated by the legend. c UMAP plots of principal identifiers of different types of myeloid cells. The intensity of expression is indicated by purple coloring. d Heatmap of top five most differentially expressed genes across myeloid clusters. The intensity of expression is indicated as specified by the color legend. e UMAP plots depicting cell identity of myeloid cells in regard to developmental time points in normally (21% O₂-exposed, purple) and aberrantly (85% O₂-exposed, green) developing lung. Each cell is colored by mouse age as indicated by the legend. f Fluorescent RNA in situ hybridization showing co-expression of Marco/MARCO (pink/red) with Ptprc/PTPRC (green/white) positive leukocytes morphologically resembling alveolar macrophages in normal and mouse hyperoxic/human BPD lungs, respectively. Magnification: 40×. Scale bar = 40 µm. Two 14-days old animals/group were analyzed and samples from five BPD patients and two donor lungs were analyzed. g Hyperoxia-impacted signaling pathways in Alv Mf (pink), Neut 1 (yellow) and Int Mf (green) 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 and violin plots represent Z-score-transformed log(TP10k + 1) values. Expression levels in UMAP plots are presented as log(TP10k + 1) values. Log(TP10k + 1) corresponds to log-transformed UMIs per 10k.