Fig. 3. EMP-derived pMacs colonize the embryo to generate macrophages.

(A) Flow cytometry analysis of E10.25 Csf1r^MeriCreMer; Rosa26^LSL-YFP (OH-TAM at E8.5) tissues showing expression of Il4ra, Il13ra1, CD16.2, CD64, Ifngr, Tnfr2, Tim4, and CD206 on YFP⁺ Kit⁺ progenitors, pMacs, and macrophages. MFI: mean fluorescent intensity. Data are representative of n=4 independent experiments with 4-6 embryos per marker. See also Fig. S3A. (B) Quantification of immunostainings on cryosection from E10.25 Csf1r^MeriCreMer; Rosa26^LSL-YFP embryos, pulse-labeled with OH-TAM at E8.5 with antibodies against YFP, Iba1 and CD16/32, Dectin-1, Trem2, F4/80, CD206 or Granulin. n=2-4 embryos and 2 sections per embryo per marker. See Fig. S4. (C) tSNE plot as in (1D) overlaid with the relative expression values for Tnfrsf11a and Cx3cr1. (D) YFP labeling efficiency of Tnfrsf11a^Cre+; Rosa26^LSL-YFP in pMacs and F4/80⁺ macrophages in YS and whole embryo at E10.25, fetal liver HSCs (long term (LT, Lin⁻Kit⁺Sca1⁺CD150⁺CD48⁻), short term (ST, Lin⁻Kit⁺Sca1⁺CD150⁻CD48⁻) and multipotent progenitor (MPP, Lin⁻Kit⁺Sca1⁺CD150⁻CD48⁺)) and tissue macrophages at E14.5 and 6 weeks, and blood leukocytes (B-cells (CD19⁺), T-cells (CD19⁻Ly6G⁻CD115⁻CD3⁺), NK cells (CD19⁻Ly6G⁻CD115⁻CD3⁻NKp46⁺), neutrophils (CD19⁻Ly6G⁺) and Ly6C^hi monocytes (CD19⁻CD115⁺Ly6G⁻Ly6C^hi), and tissue CD11b^high myeloid cells from 6 week-old mice. Circles represent individual mice. n=4 independent experiments. See Fig. S5.