Aldh3a2 depletion causes death by ferroptosis. (A) Assessment of cell cycle in Aldh3a2-control and -mutant leukemia cells reveals no differences. (B-C) Caspase 3 activation (B) and PARP cleaveage (C) in Aldh3a2-mutant vs -control LSPCs. (D-F) Frequency of live Aldh3a2-control and -mutant LSPCs 3 days after culture and treatment with vehicle (dimethyl sulfoxide [DMSO]) or the pancaspase inhibitor ZVAD (D), the ferroptosis inhibitor ferrostatin (E), or the GPX4 inhibitor RSL3 (F). (G) Sublethally irradiated C57BL/6J mice were injected with Aldh3a2-control and -mutant leukemia cells from primary leukemic mice infected with lentivirus expressing Gpx4 or scrambled shRNA. Forty-eight hours after injection, mice were injected with 3 doses of polyinosinic-polycytidylic acid [Poly(I):Poly(C)] on alternate days, and leukemia development was monitored. Kaplan-Meier survival curve of animals that developed leukemia is shown. (H) Sublethally irradiated C57BL/6J mice were injected with Aldh3a2-control and -mutant leukemia cells from primary leukemic mice. Forty-eight hours after injection, mice were treated with 3 doses of Poly(I):Poly(C) on alternate days and with cytarabine and doxorubicin in a 5 + 3 regimen. Leukemia development was monitored. Kaplan-Meier survival curve of animals that developed leukemia is shown. Data are representative of ≥2 independent experiments; n = 3 replicates per cell line per experiment. Data are represented as mean ± standard deviation. P > .05 was considered nonsignificant (ns). **P < .01, ***P < .001.