Extended Data Fig. 9: Changes in progenitor localization after infection.

a, Average percentage of the indicated cells per femur (normalized to numbers at day 0) at the indicated time points after infection of wild-type mice with L. monocytogenes. (n = 6 mice for days 0 and 2; n = 3 mice for day 4; n = 4 mice for day 6; n = 3 mice for day 8.) b, Histograms showing the distribution of distances from each MDP (green, n = 243 MDP from a total of 23 sternum sections from 15 uninfected wild-type mice and n = 37 MDPs from a total of 3 sternum sections from 3 wild-type mice 4 days after infection with L. monocytogenes), MOP (orange, n = 458 MOP from a total of 11 sternum sections from 11 uninfected wild-type mice and n = 377 MOPs from a total of 3 sternum sections from 3 wild-type mice 4 days after infection with L. monocytogenes) or GP (red, n = 338 GPs from a total of 15 sternum sections from 12 uninfected wild-type mice and n = 218 GPs from a total of 3 sternum sections from 3 wild-type mice 4 days after infection with L. monocytogenes) to the indicated progenitors 4 days after infection of wild-type mice with L. monocytogenes. c, Number of GPs or MOPs per cluster for the sternum sections analysed in b. Each dot represents a cluster; n = 17 GP clusters and n = 15 MOP clusters from 3 wild-type mice 4 days after infection with L. monocytogenes. d, Experimental design for in vivo fate map** using Ubc–cre^ERT2:confetti mice. e, Percentage of GP or MOP clusters with at least one confetti-labelled GP or MOP that are monoclonal (all cells in the cluster are labelled in the same confetti colour) or oligoclonal (containing cells with at least two different origins: CFP⁺, YFP⁺, or no confetti label). Each dot represents a GP or MOP cluster from a total of three sternum segments from two confetti mice in two experiments four days after infection with L. monocytogenes. f, Representative images showing a cluster composed of MDP-derived Gfi1^lo MOPs and GMP-derived Gfi1^hi MOPs in Gfi1–tomato mice four days after infection with L. monocytogenes. Scale bar, 10 μm. g, Percentage of GMP-derived Gfi1^hi MOPs (orange dots) or MDP-derived Gfi1^lo MOPs (purple dots) per cluster (each dot represents one cluster; n = 15 clusters in a total of 3 sternum sections from 2 Gfi1–tomato mice 4 days after infection with L. monocytogenes). h, Quantification of CD117 expression in PNs of wild-type mice at the indicated time points after infection. (n = 6 mice for day 0; n = 8 mice for days 2 and 4; n = 5 mice for day 6; n = 4 mice for day 8 in total 8 experiments). One dot indicates one mouse. i, Map showing the location of Gfi1^hi and Gfi1^lo MOPs, MDPs, and Ly6C^hi and Ly6C^lo monocytes in a sternum segment from Gfi1–tomato mice four days after infection. Scale bar, 200 μm. j, Histograms showing the distribution of distances from each Gfi1^hi (orange) and Gfi1^lo (purple) MOP and MDP (green) to the indicated cells in uninfected wild-type mice or four days after infection with L. monocytogenes. The values for day 0 (d0) are the same as shown in Fig. 2g, i. (n = 113 Gfi1^hi MOP and n = 72 Gfi1^lo MOPs from a total of 3 sternum sections from 3 uninfected Gfi1-tomato mice; n = 155 Gfi1^hi MOP and n = 144 Gfi1^lo MOP from total 3 sternum sections of 2 Gfi1–tomato mice 4 days after infection with L. monocytogenes; MDPto Ly6C^hi monocyte, n = 67 MDPs from a total of 6 sternum sections from 4 mice; MDP to Ly6C^lo monocyte, n = 67 MDPs from a total of 6 sternum sections from 4 wild-type uninfected mice; MDP to cDC, n = 139 MDPs from a total of 11 sternum sections from 6 wild-type uninfected mice; and n = 32 MDPs from a total of 3 sternum sections from 3 wild-type mice 4 days after infection with L. monocytogenes.) k, Representative image showing lack of contribution of a confetti-labelled MOP. Tracked cells are YFP⁺, CFP⁺ or unlabelled CD117⁺ CD115⁺ CD11b⁻ Ly6C⁺ MOPs, CD117⁻ CD115⁺ CD11b⁺ Ly6C^hi monocytes and CD117⁻ CD115⁺ CD11b⁺ Ly6C^lo monocytes. Scale bar, 10 μm. l, Quantification of cell numbers for CFP⁻ (white) and CFP⁺ (blue) Ly6C^hi monocytes (left) or Ly6C^lo monocytes (right) found within the indicated distances to the closest CFP⁺ MOP cell in confetti mice four days after infection with L. monocytogenes. Each dot represents one CFP⁺ MOP from a total of eight sternum segments from two confetti mice in two experiments four days after infection. m, Representative images showing the lack of contribution of a confetti-labelled MDP to surrounding monocytes. Tracked cells are YFP⁺, CFP⁺ or unlabelled CD117⁺ CD115⁺ CD11b⁻ Ly6C⁻ MDPs, CD117⁺ CD115⁺ CD11b⁻ Ly6C⁺ MOPs, CD117⁻ CD115⁺ CD11b⁺ Ly6C^hi monocytes and CD117⁻ CD115⁺ CD11b⁺ Ly6C^lo monocytes. Scale bar, 20 μm. n, qPCR analyses showing Csf1 mRNA levels (normalized to not infected) in bone marrow endothelial cells FACS-purified from wild-type mice in the steady-state or four days after infection. n = a total of three uninfected mice and n = a total of three infected mice in two experiments. o, Histogram showing the distance from each MDP to the closest sinusoid in control (pool of Cre:Csf1^+/−, Csf1^+/− and Csf1^fl/-) or Csf1^ΔEC mice four days after infection. n = 58 MDPs from a total of 4 sternum sections from 3 control mice and n = 36 MDPs from a total of 4 sternum sections from 3 Csf1^ΔEC mice. p, q, Maps (p) showing the location of the indicated cells; and histogram (q) showing the distance from each MDP to the closest Ly6C^lo monocyte and cDC in control or Csf1^ΔEC mice four days after infection. n = 51 MDPs from a total of 3 sternum sections of 3 control mice and n = 29 MDP from total 3 sternum sections of 3 Csf1^ΔEC mice. r, Number of the indicated cells per femur in control or Csf1^ΔEC mice 4 days after infection. Each dot indicates one mouse. n = 3 control and n = 3 Csf1^ΔEC mice. Unless otherwise indicated, one dot represents one cell. Statistical differences were calculated using two-tailed Student’s t-tests; P values are shown.

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