Extended Data Fig. 8: Total HSC numbers in the body remain unchanged when the size of the intact niche is increased, even in mice with impaired HSC retention in endogenous BM niches.

a, Left: representative flow cytometry plots of TdTomato⁺ cells in CD51^–CD140α^– cells and CD51⁺CD140α⁺ MSCs within CD45^–TER-119^–CD31^– fraction of Cdh2-CreER; iTdTomato mice. Right: quantification of overlap of CD51^–CD140α^–, CD51⁺CD140α⁺ and TdTomato⁺ cells in the CD45^–TER-119^–CD31^– fraction of Cdh2-CreER; iTdTomato mice. n = 4 mice. b, Left: representative flow cytometry plots of TdTomato⁺ cells in Nestin-GFP^– cells and Nestin-GFP⁺ MSCs within CD45^–TER-119^–CD31^– fraction of Cdh2-CreER; iTdTomato; Nestin-GFP mice. Right: Quantification of overlap of Nestin-GFP^–, Nestin-GFP⁺ and TdTomato⁺ cells in the CD45^–TER-119^–CD31^– fraction of Cdh2-CreER; iTdTomato; Nestin-GFP mice. n = 4 mice. c-e, HSC numbers in the femurs (c), blood (d) and spleens (e) of Cxcl12^fl/fl and Cdh2-CreER; Cxcl12^fl/fl mice. n = 7, 9 mice, respectively. f, Schematic of the transplantation of six WT femurs into Cdh2-CreER; Cxcl12^fl/fl mice and analyses. The diagram was created using BioRender. Takeishi, S. (2025) https://BioRender.com/9d3nv16. g, HSC numbers per host femur and graft of the indicated genotypes. 8 femurs from 8 sham-operated mice, 8 host femurs and 48 grafts from 8 bone transplantation hosts in both Cxcl12^fl/fl and Cdh2-CreER; Cxcl12^fl/fl groups. h-j, HSC numbers in the spleens (h), in the entire body of hosts (excluding grafts) (i) and the sum of HSCs in the hosts and the grafts (j) of the indicated genotypes. n = 8 mice per group. Data are mean ± s.e.m. Significance was assessed using a two-tailed unpaired Student’s t-test (c-e) or one-way ANOVA (g-j).

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