Fig. 9: The dysregulated hematopoiesis in aged ASXL1-MT KI mice is reverted by treatment with rapamycin.

a Aged Vav-Cre ASXL1-MT KI mice and age-matched control mice were treated with rapamycin intraperitoneally every other day for 8 weeks. At the end of administrations, end-point analyses of bone marrow cells were conducted. b, c Mitochondrial membrane potential (b) and intracellular ROS levels (c) in LT-HSCs were analyzed (n = 4 (Control-Vehicle), 5 (Control-Rapamycin), 4 (ASXL1-MT-Vehicle), and 5 (ASXL1-MT-Rapamycin)). Results are expressed as fold change relative to vehicle control group. d Absolute numbers of bone marrow cells per leg (n = 6 (Control-Vehicle), 7 (Control-Rapamycin), 5 (ASXL1-MT-Vehicle), and 7 (ASXL1-MT-Rapamycin)). e Absolute numbers of white blood cells in bone marrow per leg (n = 5 (Control-Vehicle), 6 (Control-Rapamycin), 5 (ASXL1-MT-Vehicle), and 6 (ASXL1-MT-Rapamycin)). Data are mean ± s.d. f The experimental design for transplantation using whole bone marrow cells treated with rapamycin. Whole bone marrow cells isolated from aged Vav-Cre ASXL1-MT KI mice and age-matched control mice that had been treated with rapamycin for 4 weeks were transplanted into lethally irradiated recipient mice with competitor cells. Recipient mice were continuously treated with rapamycin after transplantation. g, h 4 weeks after transplantation, effects of rapamycin treatment on mitochondrial membrane potential (g) and γ-H2AX levels (h) in LSK cells were determined (n = 4). i, j 4 weeks after transplantation, the frequency of donor-derived cells in peripheral blood and LSK cells were analyzed (n = 8) (i). Differentiation index, which is obtained by dividing the frequency of donor-derived cells in peripheral blood by LSK cells, was calculated (j). Statistical significances are assessed by one-way ANOVA with Tukey–Kramer’s post-hoc test (b–e) or two-tailed Student’s t-test (g–j). *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001.