Mortalin/DJ-1 complex formation is a key molecular mechanism underlying ROS regulation in HSCs. (A) Total number of BM cells derived from 12- and 24-week-old WT and DJ-1 knockout mice. Data represent the mean ± SD (12-week-old mice, n = 4; 24-week-old mice, n = 6). (B) The population of LSK cells (left) and CD150⁺CD48^- LSK cells (right) within the entire population of BM cells. Data represent the mean ± SD (12-week-old mice, n = 4; 24-week-old mice, n = 6; *P < .05). (C) Representative FACS profiles of LSK CD150⁺CD48⁻ fractions derived from 24-week-old WT and DJ-1 knockout mice (n = 4). (D) Representative FACS profiles (right) and the mean (±SD) DCF-DA intensity in LSK cells derived from 24-week-old WT and DJ-1 knockout mice (left) (n = 4; **P < .01). (E) Intensity of DCF-DA fluorescence in CD150⁺CD48⁻ LSK or CD48⁺ LSK cells derived from 24-week-old WT and DJ-1 knockout mice. Data represent the mean ± SD (n = 4; ***P < .001). (F) Percentage (left) and representative FACS profiles (right) of CD150⁺CD48⁻ LSK cells in the mortalin-overexpressing DJ-1^−/− and WT cell populations after 10 days of culture. Sorted mortalin-overexpressing retrovirus-transduced GFP⁺ LSK cells derived from DJ-1^−/− and control mice were used in these experiments. Data represent the mean ± SD (n = 8/group; ***P < .001). (G) Percentage (left) and representative FACS profiles (right) of Mitosox⁺CD150⁺ LSK cells in the mortalin-overexpressing DJ-1^−/− and WT cell populations after 10 days of culture. Sorted mortalin-overexpressing retrovirus-transduced GFP⁺ LSK cells derived from DJ-1^−/− and control mice were used in these experiments. Data represent the mean ± SD (n = 8/group; ***P < .001).