Extended Data Fig. 5: Interaction of myeloid progenitors with the microenvironment and with HSCs.

a, Representative images showing the simultaneous detection of PNs, INs, Ly6C^lo monocytes and cDCs. Scale bar, 10 μm. b, Map showing the location of the indicated cells in the bone marrow. Each dot corresponds to one cell. Note that the radius of each dot is two times the average radius of the cell. Scale bar, 200 μm. c, Histograms showing the distance from each Ly6C^lo monocyte (yellow dots) or cDC (pink dots) and their random simulation (white dots) to the closest indicated cell (Ly6C^lo to PN/IN, n = 500 Ly6C^lo monocytes; cDC to PN/IN, n = 727 cDCs; Ly6C^lo to cDC, n = 1,322 Ly6C^lo monocytes; from a total of 3 sternum sections from 3 mice). d, High-power images showing the relative positions of MDPs, MOPs, GPs and sinusoids. Scale bar, 10 μm. e, Histograms showing the distance from each MDP (green dots), MOP (orange dots), GP (red dots) or random distribution (white dots) to the closest indicated structure (for distances to arterioles, n = 62 MDPs from a total of 6 sternum sections of 6 mice; n = 218 MOPs and n = 114 GPs from a total of 5 sternum sections from 5 mice; for distances to endosteal surface, n = 98 MDPs, n = 410 MOPs, n = 217 GPs, from a total of 9 sterna from 6 mice). f, Representative images of multiple sternum segments, showing that MDPs are evenly distributed through the bone marrow, consistent with their sinusoidal location. Scale bars, 200 μm. g, Representative images showing the detection of HSCs and MDPs in a single stain. Scale bar, 10 μm. h, Quantification of MDP and Lin⁻ CD117⁺ CD48⁻ CD41^dim CD150⁺ HSCs in femurs by FACS (white) or imaging (orange) analyses. Each dot corresponds to one mouse femur or sternum image. n = 4. i, Representative images showing detection of HSCs and a population containing CD117⁺ Ly6C⁺ GPs and MOP in a single stain. Scale bar, 10 μm. j, Maps and histograms showing the relationships between HSCs and MDPs or GPs/MOPs in the bone marrow. In the map, the dot radius is three times the average cell radius. Scale bar, 200 μm. (n = 35 MDPs from a total of 4 sternum sections from 3 mice, and n = 191 GPs and MOPs from a total of 3 sternum sections from 3 mice.) Unless otherwise indicated, for all graphs one dot corresponds to one cell. Horizontal blue bars indicate the median distance. Statistical differences were calculated using two-tailed Student’s t-test; P values are shown.

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