Extended Data Fig. 3: The distribution of HSCs in the mouse BM is not altered after Vcam1 deletion in Csf1r-iCre⁺ cells.

(a) Representative whole⁻mount images of the sternal BM of Control and Vcam1^Csf1r-iCre mice and magnified high power view. The dashed outline denotes bone-BM border. Arterioles (Art) are identified by CD31⁺ CD144⁺ Sca1⁺ expression. Phenotypic HSCs are identified by Lineage^- CD41⁻ CD48⁻ CD150⁺ expression, and megakaryocytes (Mk) are distinguished by their size, morphology and CD41⁺ CD150⁺ expression. Representative images of n = 8 independent sternum segments (b, c) Localization of HSCs relative to (b) arterioles and (c) Mks in Vcam1^fl/fl and Vcam1^Csf1r-iCre mice. (n = 104 HSCs in Vcam1^fl/fl, n = 90 HSCs in Vcam1^Csf1r-iCre) (d) Number of BMNCs, MPP and HSC per femur in Vcam1^fl/fl and Vcam1^Csf1r-iCre mice after 5-FU injection (Vcam1^fl/fl day 0 n = 5, day 5 n = 2, day 8-20 n = 3, day 25 n = 4; Vcam1^Csf1r-iCre day 0 n = 5, day 5-15 n = 2, day 20-25 n = 4 biological replicates). (e) Spleen cellularity, and (f) number of HSCs and MPPs per spleen in Vcam1^fl/fl and Vcam1^Csf1r-iCre mice (n = 5 biological replicates). Error bars, mean ± s.e.m. Two-sample Kolmogorov–Smirnov tests were used for comparisons of distribution patterns in (b) and (c). Unpaired two-tailed student’s t test (d,e,f). Significant P values are indicated in the figure.

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