Fig. 2: Normal emergence of definitive HSCs but defective translocation to the bone marrow.

a Dot plots resolve cells of the aorta-gonad mesonephros (AGM) region of E10.5 embryos of indicated genotypes for the expression of CD45 and Kit. Cells expressing CD31 (blue) or CD27 (red) are superimposed into the dot plots (left). Quantification of Kit⁺, CD31⁺, and CD27⁺ cells per AGM (right). A two-sided unpaired Student’s t-test was conducted for statistical analysis. (n = 18, 2 biological replicates). b Plots show numbers of HSCs (KSL Slam) in the liver, spleen, and bone marrow (2 femura) of E17.5 embryos of indicated genotypes. A two-sided unpaired Student’s t-test was conducted for statistical analysis. (n = 18, 3 biological replicates for fetal liver, n = 21, 3 biological replicates for fetal spleen, n = 27, 4 biological replicates for fetal bone marrow). c Plots show the number of leukocytes (CD45), HSPCs (KSL), and HSCs (KSL Slam) in the bone marrow, spleen, and liver of newborn mice of indicated genotypes. A two-sided unpaired Student’s t-test was used. (n = 29, 4 biological replicates for bone marrow, n = 21, 4 biological replicates for spleen, n = 37, 6 biological replicates for liver). d Graphs show in vitro colony formation from bone marrow, spleen, and liver cell suspensions of newborn mice (0–4-day-old) of indicated genotypes. A two-sided unpaired Student’s t-test was used. (n = 15, 3 biological replicates for bone marrow, n = 12, 3 biological replicates for spleen, n = 14, 3 biological replicates for liver). e Graphs show the fold-change of CXCR4 mean fluorescence intensity (MFI) on HSCs in the bone marrow, spleen and liver of newborn (0–4-day-old) mice of indicated genotypes. Fold-change was calculated by dividing the individual CXCR4 MFI value of Rank^cre/+;Csf1r^fl/− HSCs to the experimental average of wild-type CXCR4 MFI values per organ. A Mann–Whitney U test was performed. (n = 8, 2 biological replicates for bone marrow, n = 13, 3 biological replicates for spleen, n = 14, 3 biological replicates for liver) f Graph shows donor cell contribution to blood neutrophils (PMN) at indicated time points after transplantation of unfractionated bone marrow cells from 2-days-old donor mice of indicated genotypes (left). Graph shows the contribution of donor-derived cells to HSCs (KSL Slam) in the bone marrow at the time point of analysis (right). A two-sided unpaired Student’s t-test was used. (n = 13, 2 biological replicates) g Plot depicts the number of CXCL12-mediated migration of liver-derived HSPCs (KSL) from 1–4-day-old mice, indicated genotypes after 2.5 h. A two-sided unpaired Student’s t-test was used. (n = 9, 3 biological replicates) h Fold-change of 1-day-old donor-derived Kit⁺ cells that have migrated to the bone marrow 16 h after transfer of whole bone marrow cells from newborn (0–1-day-old) mice of indicated genotypes into the liver of newborn wild-type recipient mice. A Mann–Whitney U test was performed. (n = 22, 2 biological replicates) Each dot represents the value for one mouse throughout the figure. Source data are provided as a Source Data file.