Generation of LT-HSCs and MPPs in a competitive environment. (A) The percentage of AKT1^−/−AKT2^−/− (CD45.2⁺Ly5^B6+) cells in the BM LSK subset, and the splenic myeloid (Mac-1⁺Gr-1⁺), B (B220⁺), and T (TCRβ⁺) lineages 6 weeks after reconstitution. Graphs represent the mean ± SEM (n = 3). (B) The gating strategy used to generate the data in panels A, C, and D. Plots are representative of one experiment 6 weeks after reconstitution. (C-D) The percentage of AKT1^−/−AKT2^−/− (CD45.2⁺Ly5^B6+) cells in the LSK, LT-HSC (CD150⁺CD48⁻LSK), or MPP (CD150^–CD48⁻LSK) populations in the bone marrow of mice at either (C) 6 weeks or (D) 12 weeks after reconstitution. Graphs represent the mean ± SEM (n = 3 for panel C and n = 5 for panel D). In panels A through D, FL cells from AKT1^−/−AKT2^−/− or littermate control animals (CD45.2⁺Ly5^B6+) were mixed with wild-type competitor FL cells (CD45.1⁺B6.Ly5^SJL) in a 1:1 ratio and injected into lethally irradiated CD45.1⁺B6.Ly5^SJL recipients. The LSK data depicted in panels A and C are the same.