Extended Data Fig. 2: The absence of Sema4A leads to functional attrition of myHSC during aging.
A. Gating strategy for quantification of overall reconstitution by donor-derived HSC and analysis of lineage composition of donor-derived graft. B. Myeloid/lymphoid ratio of peripheral blood donor-derived cells (CD45.2) in WT mice (CD45.1) which were competitively transplanted with aged WT myHSC or aged WT lyHSC (CD45.2) (n = 5 animals per group). C. Gating strategy for quantification of lineage contribution by donor-derived cells. D. Distribution of pairwise Spearman’s correlation distances between aged WT and Sema4AKO myHSC (left) and lyHSC (right) (n = 642 cells across 2 biological replicates and 4 technical replicates). In box plot, centre line shows median, box limits indicate upper and lower quartiles, whiskers extend to minimum and maximum values E. Volcano plots showing DEGs in myHSC from aged WT and Sema4AKO mice. The x and y axes indicate the expression fold change (FC) (log2) and the false discovery rate (FDR) ( − log10) for each gene versus controls, respectively. Legends highlight upregulated (red) or downregulated (blue) transcripts, as well as genes not passing cutoff criteria for FC (black) and FDR (gray). Selected representative genes are shown (n = 2 biological replicates per genotype). F. Distribution of diffusion pseudotime values of myHSC and lyHSC from aged WT mice (n = 642 cells across 2 biological replicates and 4 technical replicates). In the inner box plots of the violinplots, the white point shows median value, box limits indicate upper and lower quartiles, whiskers extend to minimum and maximum values. P values are shown. Statistical significance was assessed by two-tailed t-test (*P < 0.05, **P < 0.01), except for the diffusion pseudotime analysis where two-tailed Wilcoxon rank sum test was used. Mean +/- SEM are shown.
