Extended Data Fig. 4: Immunomodulatory roles of the GYPA+CD71+CD63+ cells in UCB.
a, The regulatory network of the selected regulons and their targets in immune-erythroid cluster. b, Enriched KEGG pathways of the target genes in (a). P values: hypergeometric test and Benjamini–Hochberg method for multiple testing. c, Enriched GO terms in the ‘Classical-Ery’ and ‘Immune-Ery’ clusters from Fig. 4k. P values: hypergeometric test and Benjamini–Hochberg method for multiple testing. d, Flow cytometry plots showing GYPA+CD71−CD63+ enucleated erythrocytes from UCB. New methylene blue staining of GYPA+CD71− cells; scale bar, 10 mm. Bar graph showing the percentage of CD63+ in GYPA+CD71− cells in UCB (n = 3 samples). Data = mean ± SEM. e, Differentially expressed immune signature genes between two groups. f, Flow cytometry plot depicting GYPA+CD71+ erythroid cells at day 14 of differentiation. Pie chart showing the composition of erythroid precursors. Wright-Giemsa staining showing their morphology (n = 3 samples); scale bar, 20 μm. g, t-SNE visualization of clusters in day 14 differentiated erythroid precursors. h, FeaturePlots showing the expression of indicated genes. i, Schematic diagram showing the experimental design. j, Heatmap showing the relative expression of immune-related genes in PBMCs with or without LPS stimulation by low-input RT-qPCR. k, Boxplot showing the expression of key immune-related genes (listed in j) in each indicated group (n = 3 independent experiments). The horizontal line indicates the median value; the box represents the first and third quartiles. P values (left, 0.034; right, 0.016): two-sided Wilcoxon rank-sum test. l, Cytokines secreted by PBMCs treated with or without LPS (n = 2 independent experiments). m, Flow cytometry plots illustrating CD71+TO+CD63+ and CD71+TO+CD63− reticulocytes from CD71 enriched, Hoechst− UCB MNCs. New methylene blue staining and Wright-Giemsa staining of CD71+TO+CD63+ and CD71+TO+CD63− reticulocytes (n = 3 samples); scale bars, 10 μm. n, Cytokine production from the indicated groups (n = 2 independent experiments). o, Schematic diagram showing the experimental design. p, Flow cytometry plots illustrating the TNF expression in the indicated groups. q, Bar graph indicating the percentage of TNF expressing cells in indicated groups (n = 5 independent experiments). Data = mean ± SEM. P values (P < 0.0001 for both): unpaired one-way ANOVA. * P < 0.05; *** P < 0.001.
