Fig. 5: Ahed plays essential roles in adult haematopoiesis.

a Schematic of the procedure. Tamoxifen injection to Rosa26-CreERT2 Ahed^fl/+ (R26CE; control) and Rosa26-CreERT2 Ahed^fl/fl (R26CE Ahed^fl/fl; cKO) mice for 5 consecutive days and analysis of BM on day 10. b Hematoxylin and eosin (H&E) staining of bone marrow tissues from R26CE and R26CE Ahed^fl/fl mice. Scale bars, 100 μm. Results shown are representative of two independent experiments. c Peripheral blood counts; n = 3, in R26CE (control) and n = 6, in R26CE Ahed^fl/fl (cKO). d A total of 3.0 × 10² LSK cells from BM were subjected to methylcellulose culture for 9 days. The numbers of colonies are shown in a right graph. Scale bars, 1 cm. e Flow cytometry plots of E14.5 foetal liver erythrocytes from R26CE and R26CE Ahed^fl/fl mice to evaluate erythroid differentiation using TER119 and CD71. Subset groups were defined as follows; S0, TER119⁻CD71⁻; S1, TER119⁻CD71⁺; S2, TER119^LoCD71⁺; S3, TER119⁺CD71⁺; S4, TER119⁻CD71⁺. Numbers indicate the percentage of each fraction. f Transplantation strategy. Donor Ahed^fl/fl (control) or R26CE Ahed^fl/fl (cKO) BM LSK cells sorted from CD45.1⁻CD45.2⁺ and transplanted into lethally irradiated CD45.1⁺CD45.2⁻ WT mice (n = 4 for both group) with CD45.1⁺CD45.2⁻ WT BM cells as a rescue. g Chimerism of peripheral blood in recipients. Numbers in each figure indicate percentages of each fraction. h Transplantation strategy. Donor Ahed^fl/fl (control) or R26CE Ahed^fl/fl (cKO) BM LSK cells were sorted by FACS from CD45.1⁻CD45.2⁺ BM mononuclear cells treated with 5 μM 4-OHT for 54 hours and transplanted into lethally irradiated CD45.1⁺CD45.2⁻ WT mice (n = 4 for control and n = 8 for cKO group). i Chimerism of peripheral blood in recipients. Numbers in each figure indicate percentages of each fraction. Data are presented as mean ± s.d. Statistical significance in c was determined by two-sided unpaired Student’s t-test, in d by one-way ANOVA with the Tukey–Kramer post-hoc test, and in e, g, i by one-way ANOVA with Brown-Forsythe test. *p < 0.05; **p < 0.01, ***p < 0.001, ****p < 0.0001. Source data are provided as a Source Data file.