Novel ALDH2 mutant are sensitive to ethanol toxicity. (A) Cellular ATP levels were measured in transiently transfected 3T3 cells using ATP-based CellTiter-Glo Luminescent Cell Viability kit. n = 8; Mean, standard deviation, probability by one‐way ANOVA (with Fischer's LSD post hoc test) ** p-value <0.01; * p-value <0.05. (B) Total cellular ROS levels were measured with 2,7 dichloro- fluorescein diacetate in cells treated as in (A). n = 8; Mean, standard deviation, probability by one‐way ANOVA (with Fischer's LSD post hoc test) ** p-value <0.01; * p-value <0.05. (C) Lactate dehydrogenase activity levels in supernatant in cells treated as in (A). n = 8; Mean, standard deviation, probability by one‐way ANOVA (with Fischer's LSD post hoc test) ** p-value <0.01; * p-value <0.05. (D) Cellular ATP levels were measured in transiently transfected 3T3 cells in the presence or absence of Alda-1/Alda-64 (20 μM/48 h; 50 mM Ethanol) using ATP-based CellTiter-Glo Luminescent Cell Viability kit. n = 8; Mean, standard deviation, probability by one‐way ANOVA (with Fischer's LSD post hoc test) **** p-value <0.0001; ** p-value <0.01; * p-value <0.05. (E) Total cellular ROS levels were measured with 2,7 dichloro- fluorescein diacetate in 3T3 cells treated as in (D). n = 8; Mean, standard deviation, probability by one‐way ANOVA (with Fischer's LSD post hoc test) **** p-value <0.0001; ** p-value <0.01; * p-value <0.05. (F) Lactate dehydrogenase activity levels in supernatant in 3T3 cells treated as in (D). n = 8; Mean, standard deviation, probability by one‐way ANOVA (with Fischer's LSD post hoc test) **** p-value <0.0001; ** p-value <0.01; * p-value <0.05. (G) 4-HNE protein adducts in transiently transfected human-derived fibroblasts treated as in (D). n = 3; Mean, standard deviation, probability by one‐way ANOVA (with Fischer's LSD post hoc test) **** p-value <0.0001; *** p-value <0.001; ** p-value <0.01; * p-value <0.05.