Figure 3. Heterozygous deletion of Kdm6b restores ASXL1^aa1–587–mediated HSC phenotypes and myeloid differentiation.

(A) Flow cytometric analysis of myeloid cells in PB from representative mice of each genotype. (B) The frequencies of Gr1⁺Mac1⁺ cells in PB from WT (n = 19), Asxl1^Y588XTg (n = 19), Kdm6b^Δ/+ (n = 15), and Asxl1^Y588XTg Kdm6b^Δ/+ (n = 15) mice. (C and E) Flow cytometric analysis of HSPCs in BM cells from representative mice of each genotype. (D and F) Quantification of the percentages of LSK cells (D) and LT-HSCs (F) in BM from WT (n = 19), Asxl1^Y588XTg (n = 19), Kdm6b^Δ/+ (n = 15), and Asxl1^Y588XTg Kdm6b^Δ/+ (n = 15) mice. (G and H) Colony-forming assay using BM cells from WT, Asxl1^Y588XTg, Kdm6b^Δ/+, and Asxl1^Y588XTg Kdm6b^Δ/+ mice (n = 10 per genotype). (I) Serial cell replating assays using whole BM cells (n = 3 mice per genotype) were performed to determine HSC self-renewal capability. The cells were replated weekly for 4 weeks. (J) Percentages of donor-derived CD45.2⁺ cells in the PB of recipient animals at indicated time points (n = 5 per genotype). Data represent the mean ± SEM. **P < 0.01 and ***P < 0.001 vs. WT mice, and ^#P < 0.05, ^##P < 0.01, and ^###P < 0.001 vs. Asxl1^Y588XTg mice, by 1-way ANOVA with Tukey’s multiple-comparison test (B, D, and F–J).