Extended Data Fig. 2: Generation and characterization of HlfCreERT2 mice.
From: Independent origins of fetal liver haematopoietic stem and progenitor cells
a, Experimental design for precursor labeling by CreERT2 lines. 1; Labeling of endothelial cells (e.g. Runx1-CreERT2 and VEcad-CreERT2). Labeled endothelial cells may transform into hematopoietic cells at a later time, leading to the mixture of cells at various stage of maturation. 2; Labeling of hematopoietic clusters. EHT, endothelial-to-hematopoietic transition. b, Whole-mount immunostaining analysis of E10.5 (35 sp) HlftdTomato embryos for tdTomato (red), c-Kit (green) and CD31 (white). Scale bar, 100 μm. c, Whole-mount immunostaining analysis of E10.5 (33 sp) HlftdTomato yolk sac for tdTomato (red), c-Kit (green) and CD31 (white). Scale bar, 50 μm. d, Targeting strategy of the HlfCreERT2 mouse. e, White blood cell (WBC), red blood cell (RBC), and platelet (PLT) fractions in the blood of adult Hlf+/+ and HlfCreERT2/+ mice (Hlf+/+ mice, n = 7; HlfCreERT2/+ mice, n = 9). f, Bone marrow (BM) cellularity (Hlf+/+ mice, n = 5; HlfCreERT2/+ mice, n = 6). g, HSC and progenitor fractions in HlfCreERT2/+ mice (Hlf+/+ mice, n = 4; HlfCreERT2/+ mice, n = 5). h, Whole-mount immunostaining analysis of E10.75 (tamoxifen at E10.0) HlfCreERT2ROSAtdTomato embryos. Scale bars, 50 μm. i, Frequency of tdTomato+ cells in c-Kit+ cells (Dorsal aorta, n = 6; Yolk sac, n = 3; Vitelline artery, n = 6; Umbilical artery, n = 6). j, HSC analysis. HlfCreERT2ROSAtdTomato embryos were administered with tamoxifen at E10.5 and were analyzed at E14.5. k, EMP analysis. HlfCreERT2ROSAtdTomato embryos were administered with tamoxifen at E8.0 and were analyzed at E10.5. l, Contribution of HlfCreEERT2-labeled cells to adult hematopoiesis. HlfCreERT2ROSAtdTomato embryos were administered with tamoxifen at E9.75. n = 5. m, Lympho-myeloid progenitor (LMP) analysis. HlfCreERT2ROSAtdTomato embryos were administered with tamoxifen at E9.75 and were analyzed at E11.5. n = 8. All error bars represent means ± SD. Statistical analysis was performed using two-sided unpaired Student’s t-test (e–g).
