Fig. 7: ASXL1-MT induces mitochondrial activation, overproduction of ROS, and increased DNA damage.

a Gene set enrichment analysis of TCA cycle and respiratory electron transport gene sets in LSK cells of young control and Vav-Cre ASXL1-MT KI mice (n = 3). b Mitochondrial membrane potential in LK cells, LSK cells, MPPs, ST-HSCs and LT-HSCs of young control and Vav-Cre ASXL1-MT KI mice (n = 7). Representative histogram in LSK cells is shown (left panel). Summarized results are expressed as fold change relative to their respective counterparts in littermate controls (right panel). c Oxygen consumption rates (OCR) in pooled 3 × 10⁵ Lin⁻c-kit⁺ cells under basal conditions and in response to oligomycin, fluoro-carbonyl cyanide phenylhydrazone (FCCP) and antimycinA/rotenone. Cells were obtained from three animals per genotype. Data are mean ± s.e.m. d Representative immunofluorescence for Tom20 in MPPs and LT-HSCs (scale bars, 5 μm). e Quantification of mitochondrial fragment (left panel), area (middle panel), and area normalized to one fragment (right panel) (30 cells/group). Cells were obtained from 2 animals per genotype. Data are mean ± s.e.m. f Intracellular ROS levels in LK cells, LSK cells, MPPs, ST-HSCs, and LT-HSCs of young control and Vav-Cre ASXL1-MT KI mice (n = 7). Representative histogram in LSK cells is shown (left panel). Summarized results are expressed as fold change relative to their respective counterparts in littermate controls (right panel). g Comet analyses using LT-HSCs from young and aged Vav-Cre ASXL1-MT KI mice (n = 3). Representative images (scale bars, 20 μm) (left panel) and olive tall moment (right panel) are shown. Statistical significances are assessed by two-tailed Mann–Whitney’s u-test (b, f), two-tailed Student’s t-test (c,s e), or one-way ANOVA with Tukey–Kramer’s post-hoc test (g). *P ≤ 0.05, **P ≤ 0.01.