Functional identification of Dnmt3a^+/−Flt3^ITD LSC population. (A-D) Moribund de novo Dnmt3a^+/−Flt3^ITD AML (n = 4) and age-matched Flt3^ITD MPN (n = 5) and WT control (n = 4) mice. (A) Average ±SEM total BM. (B) Average ± SEM HSC (CD150⁺CD48⁻CD34⁻CD135⁻LSK), MPP1, MPP2, and MPP3 cells per million total BM. (C) Average ± SEM total spleen. (D) Average ± SEM HSC (CD150⁺CD48⁻CD34⁻CD135⁻LSK), MPP1, MPP2, and MPP3 cells per million total spleen. (E) Experimental schematic for panels F,G. Immunophenotypic HSC (CD150⁺CD48⁻CD34⁻CD135⁻LSK), MPP1 (CD150⁺CD48⁻CD34⁺CD135⁻LSK), or MPP2 (CD150⁺CD48⁺CD34⁺CD135⁻LSK) Dnmt3a^+/−Flt3^ITD AML cells (CD45.2⁺) were sorted and transplanted into CD45.1⁺ WT recipient mice. Peripheral blood (PB) chimerism evaluated at indicated time points after transplant. (F) Average ±SEM percent CD45.2⁺ cells in PB (n = 4 HSC/MPP1/MPP2 matched donor sets with n = 1 recipient/cell-type). (G) LSC estimation for each Dnmt3a^+/−Flt3^ITD AML cell-type from panel F. Statistical significance by ordinary 1-way ANOVA using Tukey multiple comparison test (A-D), or ELDA analysis (G).