Extended Data Fig. 8: CSF1 from a subset of endothelial cells is necessary for the generation of dendritic cells.

a, qPCR analyses showing Csf1 mRNA levels (normalized to endothelial control, Ctrl) in FACS-purified endothelial cells and the indicated haematopoietic cells in control or Csf1^ΔEC mice; n = a total of four control and n = a total of four Csf1^ΔEC mice in four experiments. Note that although Cdh5–cre also recombines in subsets of haematopoietic cells, it does not affect Csf1 expression in these cells. (n.d., none detected; n = a total of three control and n = a total of three Csf1^ΔEC mice in three experiments.) EOS, eosinophil; Ery, erythrocyte. b, Colony-forming activity (blue, GMPs; green, MDPs; orange, MOP; red, GPs) of the indicated progenitors FACS-purified from control (diamonds, n = 5) or Csf1^ΔEC (circles, n = 5) mice in four experiments. c, d, Number (c) and CFU-M activity (d) of GMPs or MOPs from control or Csf1^ΔEC mice. n = a total of six control and n = a total of six Csf1^ΔEC mice in five experiments. e, Number of Ly6C^hi and Ly6C^lo monocytes in the bone marrow and peripheral blood of control or Csf1^ΔEC mice. n = a total of eight control and n = a total of eight Csf1^ΔEC mice in six experiments. f–h, Number of bone marrow cells from the indicated populations in the femur or blood of control or Csf1^ΔEC mice. n = a total of eight control and n = a total of eight Csf1^ΔEC mice in six experiments. i, Volume and number of vessels in sternum sections of control or Csf1^ΔEC mice. Each dot represents one sternum segment, from three control and three Csf1^ΔEC mice. j, Percentages of the indicated CD45.2⁺ cells in the blood of lethally irradiated CD45.1⁺ recipients after transplant of 10⁶ bone marrow cells purified from control (white dots) or Csf1^ΔEC (red dots) mice—both CD45.2⁺—together with 10⁶ CD45.1⁺ competitor cells at the indicated time points after transplantation. Shown are means ± s.d. from 12 mice per group. k, Representative images showing anti-CSF1 or isotype control stains in the bone marrow of wild-type mice. Scale bars, 200 μm and 10 μm. l, Map of CSF1⁺ and CSF1⁻ vessels and cDCs in Csf1^ΔEC mice. Scale bar, 200 μm. m, High-power images showing a CSF1⁺ vessel and cDCs (pink dots) in control mice. The radius of the dot is two times the average cDC radius. Scale bar, 20 μm. n, Number of cDCs found within the indicated distances of CSF1⁺ and CSF1⁻ vessels in wild-type (n = 76 CSF1⁺ vessels and n = 520 CSF1⁻ vessels in a total of 4 sternum sections from 3 wild-type mice). o, Histograms showing the distance from each cDC to the closest sinusoid in control or Csf1^ΔEC mice (n = 451 cDCs in a total of 2 sternum sections from 2 control mice; n = 343 cDCs in a total of 3 sternum sections from 3 Csf1^ΔEC mice). p, Maps showing the relocation of MDPs away from sinusoids in Csf1^ΔEC mice. Scale bars, 200 μm and 10 μm. The radius of the dots is three times (left) or one times (right) the average radius of the MDP. q, Maps showing the distribution of MDPs, Ly6C^lo monocytes and cDCs in the sternum of control or Csf1^ΔEC mice. Scale bar, 200 μm. The radius of the dot is three times (for MDPs) or two times (for all other cells) the average radius of the replaced cell. r, Histograms showing the distribution of distances from each MDP to the six closest Ly6C^lo monocytes or cDCs in control or Csf1^ΔEC mice. (For MDPs to Ly6C^lo monocytes, n = 37 MDPs from a total of 4 sternum sections from 3 control mice; n = 18 MDPs from a total of 4 sternum sections from 3 Csf1^ΔEC mice. For MDPs to cDCs, n = 47 MDPs from a total of 6 sternum sections from 3 control mice; n = 47 MDPs from a total of 9 sternum sections from 3 Csf1^ΔEC mice.) Unless otherwise indicated, for a–i each dot corresponds to one mouse; for l–r each dot corresponds to one cell. Statistical differences were calculated using two-tailed Student’s t-tests; P values are shown.

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