Aldh3a2 depletion alters the redox state of cells. (A-B) Levels of endogenous 16- and 18-carbon alcohols (A) as well as total alcohol levels (B) in Aldh3a2-control and -mutant leukemic cells. (C-D) Cellular ROS (C) and lipid peroxidation (D) levels in Aldh3a2-control and -mutant LSPCs (Lin^lowc-Kit⁺). (E-F) Cellular ROS (E) and lipid peroxidation levels (F) in Aldh3a2-control and -mutant LSPCs upon 4-HNE and 4-HNE plus vitamin E treatment. (G) Growth kinetics of Aldh3a2-control and -mutant LSPCs treated with 4-HNE. (H) Growth kinetics of Aldh3a2-control and -mutant N-GMPs treated with 4-HNE. (I-K) Aldh3a2-mutant leukemia cells show evidence of oxidative damage to DNA and protein as shown by increased levels of γ-H2AX (double-stranded DNA breaks) (I), 8-OHDG (oxidative DNA damage) (J), and protein carbonylation (oxidative protein damage) (K) in mutant vs control cells. Data are representative of ≥2 independent experiments; n = 3 replicates per cell line per experiment (except for measurement of alcohols, where 2 replicates were used). Data are represented as mean ± standard deviation. P > .05 was considered nonsignificant (ns). *P < .05, **P < .01, ***P < .001. DMSO, dimethyl sulfoxide; FITC, fluorescein isothiocyanate; MFI, mean fluorescence intensity.