Supplementary Figure 6: In vivo myeloid reconstitution by GE− and GE+ myeloid progenitor cells.
From: Distinct myeloid progenitor–differentiation pathways identified through single-cell RNA sequencing
(a) Experimental design for direct comparison of in vivo potential from GE− and GE+ GMPs and pre-GMs. (b-c) Quantification of in vivo contribution of GE+ and GE− GMP (b) and pre-GM (c) progenitors to mast cells, eosinophils, neutrophils and monocytes, 7 (red; n=3) and 10 (blue; n=5) days after intra-femur transplantation of 2000-11000 cells. Mean (SD) donor derived cells per million analyzed MNCs per 1000 injected cells detected in injected femur (top) and spleen (bottom) is shown. Data is from three independent experiments. * below detection limit (see methods). (d) Total donor (CD45.2 + CD45.1/2) contribution per million MNCs analyzed at 7 and 10-11 days following IP or IF transplantation of indicated progenitor populations. Peritoneal fluid was analyzed from IP transplanted mice, and bone marrow cells from injected femur and spleen cells were analyzed from IF transplanted mice (n=3-5 mice). 1000-4000 cells/mouse were injected IP and 2000-11000 cells/mouse were injected IF. Data is adjusted to per 1000 injected donor cells per 1 million acquired MNCs. (e-h) Representative morphology of donor-derived monocytes (e), neutrophils (f), eosinophils (g) and mast cells (h) following IP transplantation of purified GMPs, pre-GMs and LMPPs. (i) Gata1-EGFP expression in donor-derived (CD45.1/2 and CD45.2) mast cells, eosinophils, neutrophils and monocytes 7 and 11 days following I.P. transplantation of GMPs and pre-GMs respectively. Data from transplanted mice included in Figure 8 is shown.
