Extended Data Fig. 6: Isolation and clustering of terminally differentiated erythroid precursors from adult BM.
a, Representative flow cytometry plots showing the gating strategy for isolation of erythroid precursors from adult BM (n = 7 samples). b, UMAP plots of each individual sample from BM. c, Hierarchical clustering of GYPA+ erythroid precursors from YS, FL, PT-UCB, term UCB and adult BM. d, e, Boxplot showing the number of genes captured (d) and erythroid maturation score (e) in BM erythroid precursors (C1, n = 578 cells; C2, n = 302 cells; C3, n = 771 cells; C4, n = 553 cells). The horizontal line indicates the median value; the box represents the first and third quartiles. f, Heatmap showing the relative expression (scaled by row) of the top 10 signature genes of each indicated cluster. g, Boxplots showing the enriched scores of the activation of immune responses and cytokine-mediated signaling pathway in the four clusters (C1, n = 578 cells; C2, n = 302 cells; C3, n = 771 cells; C4, n = 553 cells). The horizontal line indicates the median level; the box represents the first and third quartiles. P values were determined by two-sided Wilcoxon rank-sum test (all P values < 10−16). h, Representative flow cytometry plots illustrating the TNF expression. The upper panel showing the TNF expression in BM-derived GYPA+CD71+ and GYPA+CD71+CD63− cells cultured alone. The lower panel showing the TNF expression in CD11b+ cells from groups of CD11b+ cells cultured alone, co-cultured with BM-derived GYPA+CD71+CD63+ cells or GYPA+CD71+CD63− cells. i, Bar graph indicating the percentage of TNF expressing cells in each indicated group from Extended Data Fig. 6h. Each dot indicates an independent experiment (n = 5 independent experiments). Data are presented as the mean ± SEM. P values were determined by unpaired one-way ANOVA (P = 0.0373, CD11b+ vs. CD11b+ + GYPA+CD71+CD63+; P = 0.0008, CD11b+ + GYPA+CD71+CD63− vs. CD11b+ + GYPA+CD71+CD63+). * P < 0.05, *** P < 0.001. j, Representative GO terms enriched from the common and developmental stage-specific signature genes in immune-erythroid cells in Fig. 5k. P values were determined by hypergeometric test and adjusted for multiple testing using the Benjamini–Hochberg method.
