Depletion of CHD8 causes a drastic loss of HSPCs. (A) Flow cytometry of LSK and LK cells from WT and Chd8^−/− mice after 6 days of pI/pC injection. Representative dot plots and frequencies of the HSPC subpopulations in lineage⁻ cells are shown. (B) WT indicates Chd8^F/F mice. Absolute number of LSK and LK cells from WT and Chd8^−/− mice after 6 days of pI/pC injections. ***P < .001. (C) Number of colonies after 7-day plating. Total BM cells were harvested 6 days after pI/pC injection and plated in MethoCult medium. ***P < .001. (D) Immunophenotypic analysis of SLAM HSC (LSK CD150⁺CD48⁻), MPP2 (LSK CD150⁺CD48⁺), and MPP3 (LSK CD150⁻CD48⁺) cells from WT and Chd8^−/− BM. Averaged frequencies of HSPC subpopulations in LSKs are shown. (E) Absolute number of SLAM HSC, MPP2, and MPP3 cells from WT and Chd8^−/− mice. **P < .01; ***P < .001. (F) The experimental scheme of the native competitive transplant into CD45.1 WT recipient. (G) Percentage of CD45.2 cells at different time points in the PB after pI/pC injection in the successfully reconstituted mice. Statistical analysis was performed at 16 weeks after deletion. ***P < .001. (H) Percentage of donor-derived mature lineage cells after 4 months of deletion. ***P < .001. (I) Percentage of CD45.2 BM progenitors in the transplant-recipient mice after 4 months of pI/pC injections. **P < .01; ***P < .001. ns, not significant. (J) The proportion of CD45.2-derived cells in PB in the secondary and tertiary transplants. Statistical analysis was performed at 16 weeks after transplantation. ***P < .001. (K) The proportion of CD45.2-derived BM progenitors at 4 months in the secondary and tertiary transplants. ***P < .001.