Extended Data Fig. 6: Molecular and functional characterization of Selp^KO HSCs.

(a) Validation of Selp deletion⁵² at exon 3 in the Selp^KO vs Selp^WT HSC RNA-seq data. The coverage of the alignments at Selp exon 3 was visualized by IGV. (b-c) Correlation analyses of genes upregulated in Selp^KO HSCs with upregulated genes in HSC aging, IL6 activating genes and IL-1β activating genes using single-cell expression data from GSE70657 ref. ⁴⁰ (b) and GSE59114 ref. ⁴¹ (c), respectively. Values on x and y axis are the enrichment scores of the gene sets in single cell. Correlation analyses using Pearson coefficient R and linear regression t-test. Gene sets can be found in Supplementary Tables 1 and 4. Linear regression line and 95% confidence interval is presented. (d) WBC, RBC and PLT counts for Selp^WT and Selp^KO mouse PB (n = 4). Data are presented as mean values +/− SD. (e) Validation of P-selectin presentation by FACS in Selp^WT and Selp^KO BM. (f) Frequencies of CMP, GMP and MEP in Selp^WT and Selp^KO BM (n = 4). **P = 0.002 (CMP), ***P = 0.0002 (GMP), *P = 0.027 (MEP), two-tailed t-test. Data are presented as mean values +/− SD. (g) Frequencies of DCs, monocytes (Mon), neutrophils (Neu) macrophages (Mac) CD8 T cells CD4 T cells and B cells in Selp^WT and Selp^KO BM (n = 4). *P = 0.027, two-tailed t-test. Data are presented as mean values +/− SD. (h) Frequencies of 50 Selp^WT and Selp^KO HSCs-derived BM cells in primary recipients. Data are from 2 independent replicates (n = 12). P = 0.0810 (Paired t-test). (i) Transplantation design of 200 Selp^WT and 200 Selp^KO LT-HSCs (n = 3). (j) WBC chimerism in PB of transplantation of 200 Selp^WT and Selp^KO LT-HSCs at indicated weeks (n = 3). **P = 0.0071 (week 8), ***P = 0.0006 (week 12), ***P = 0.0003 (week 16), two-way ANOVA followed by Sidak test. Data are presented as mean values +/− SD. (k) Frequencies of Myeloid, B and T cells in PB of 200 Selp^WT and 200 Selp^KO HSC transplantation at indicated weeks (n = 3). Data are presented as mean values +/− SD.

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