Quantification of AKT1^−/− or AKT2^−/− HSCs in adult mice and assessment of function in serial transplantation. Total number of LSKs and LT-HSCs (CD150⁺CD48⁻LSKs) in bone marrow from femurs and tibias in (A) 3-month-old or (B) 12-month-old single knockout mice. Graphs represent the mean ± SEM in (A) n = 6 and (B) n = 4. (C) Percentage of single knockout (CD45.2⁺Ly5^B6+) cells in the bone marrow LSK population 16 weeks after the initial transplantation (primary) and then 16 weeks after each serial transplantation (secondary and tertiary). FL cells from single-knockout fetuses (CD45.2⁺Ly5^B6+) were mixed with wild-type FL cells (CD45.1⁺ B6.Ly5^SJL) in a 3:1 (CD45.2⁺:CD45.1⁺) ratio and serially transplanted every 16 weeks. Bar graphs represent the mean ± SEM (n = 4).