Altered gene expression profiles in HSCs derived from physioxia and ambient air exposure as assessed by CITE-seq analysis. Three biological replicates are analyzed. (A) CITE-seq and cell hashing workflow under physioxia and ambient air. (B) UMAP of CITE-seq data colored by computationally predicted cell clusters. LSK cell types can be discerned by their surface protein marker expression profiles: (1) CD150-CD117-air, (2) CD150-CD117-physioxia, (3) CD150-CD117-physioxia, (4) CD150-CD117-CD48-physioxia, (5) CD150-CD117-CD48-air, (6) CD150 high-physioxia, (7) CD150 high-air, (8) CD150 very high-physioxia, (9) CD150 very high-air, (10) CD48 high-physioxia, (11) CD48 high-air, (12) CD48 mid-air. (C) UMAP of WT BM CD150^hi cells under physioxia and ambient air. (D) Heatmap of averaged expression level of HSC renewal and marker genes in WT CD150^hi cells under physioxia and ambient air. (E) GSEA plots of selected differentially expressed pathways in WT CD150^hi cells under physioxia and ambient air. (F) Heatmap of averaged expression level and GSEA of genes in the p38 MAPK pathway in WT CD150^hi cells under physioxia and ambient air. (G) The violin plots show the normalized expression of Tet2 in WT LSK CD150^hi cells under physioxia and ambient air. (H) 5-hmC levels in WT BM Lin⁻ cells under physioxia and ambient air; data are presented as mean ± standard error of the mean. ****P < .0001 when analyze by Wilcoxon rank sum test (G) and Student t test (H). KEGG, Kyoto Encyclopedia of Genes and Genomes; PID, The Pathway Interaction Database; UMAP, Uniform Manifold Approximation and Projection.