Fig. 4: Ahed is indispensable for HSPC differentiation.

a Absolute numbers of LSK cells in E14.5 foetal liver (FL). b Absolute numbers of LT-HSCs, MPPs, LMPPs, and CLPs (n = 3–5), pooled from two independent experiments. In bar charts, results are shown as mean ± s.d. and each dot represents an individual FL result. c Absolute numbers of CMPs, GMPs, MEPs (n = 2–6). d A schematic representation summarizing (a–c). e LSK cells from E14.5 FL were subjected to methylcellulose culture. Generated colonies were classified and counted. BFU-E, burst-forming units-erythroid; CFU-G/M/GM, colony-forming unit-granulocyte/macrophage/granulocyte-macrophage. f E14.5 FL LSK cells were co-cultured with MS-5 in the presence of SCF, Flt3-ligand and IL-7. Flow cytometry profiles of recovered CD45⁺ cells (left). Absolute numbers of CD19⁺ B cells differentiated from 2.5 × 10² LSK cells (right). g Limiting dilution analysis of E14.5 FL LSK cells using MS-5 co-culture. Frequencies of haematopoietic progenitor cells were determined and shown in parenthesis. NA, not analysed. h Transplantation strategy. Donor Vav1-cre Ahed^fl/+ (control) or Vav1-cre Ahed^fl/fl (cKO) E14.5 LSK cells sorted from CD45.1⁻CD45.2⁺ and transplanted into lethally irradiated CD45.1⁺CD45.2⁺ WT mice (n = 7–8) with 5 × 10⁵ CD45.1⁺CD45.2⁺ WT BM cells as a rescue. i Chimerism of peripheral blood in recipients. Numbers in each figure indicate percentages of each fraction. Results shown are representative of two independent experiments. j Chimerism of LSK in recipients’ bone marrow. Numbers in each figure indicate percentages of each fraction. Results shown are representative of two independent experiments. Data are presented as mean ± s.d. Statistical significance in a-c, i, j was determined by one-way ANOVA with the Tukey–Kramer post-hoc test, and in e, f by one-way ANOVA with Brown-Forsythe test. *p < 0.05; **p < 0.01; **p < 0.001; ****p < 0.0001. NS, not significant. Source data are provided as a Source Data file.