Extended Data Fig. 2 |. ALDH2 expression in different brain regions of Aldh2^Gfap−/− mice.

a, the schematic diagram shows the generation of astrocytic specific Aldh2 deficient mice. b, A representative image of PCR genotyping result. The image of agar gel migration shows flox (homozygous: one band, 365 bp; heterozygous: two bands, 365 and 300 bp) and Cre (negative: one band, 350 bp; positive: two bands, 350 and 200 bp) gene. c, RNAscope images of ALDH2 signals in cerebellum and liver slices from conditional and global Aldh2^−/− mice. Noted that ALDH2 was selectively depleted from microdomains surrounding PC signal in cerebellar slice from Aldh2^Gfap−/− mice. Liver ALDH2 mRNA was intact in Aldh2^Gfap−/− mice, whereas liver ALDH2 signal was abolished in Aldh2^−/− mice. Each experiment was repeated four times with similar results obtained. White scale bars, 100 μm. d, e, Representative peaks from electrophoresis densitometry of cerebellar (n=7, 7 and 3 mice, respectively) and liver (n=5, 5 and 3 mice, respectively) ALDH2 proteins from Aldh2^Gfap+/+ (black line or rectangle), Aldh2^Gfap−/− and Aldh2^−/− mice. f, Summary statistics of cerebellar ALDH2 enzymatic activity detected in Aldh2^L7−/− and Aldh2^{Camk2a−/−} mice (n=6, 6, 7 and 7 mice, respectively). g, Representative electropherogram traces of ALDH2 protein samples collected from different brain areas of Aldh2^Gfap+/+ and Aldh2^Gfap−/− mice. h, Summary statistics of brain ALDH2 protein detected in difference brain areas from Aldh2^Gfap+/+ (n=5, 4, 6, 4, 4 and 7 mice, respectively) and Aldh2^Gfap−/− (n=5, 4, 6, 4, 4 and 7 mice, respectively) mice. i, Representative electropherogram traces of protein samples collected from different brain areas from Aldh2^Camk2a+/+ and Aldh2^{Camk2a−/−} mice. j, Summary statistics of brain ALDH2 protein detected in difference brain areas from Aldh2^Camk2a+/+ and Aldh2^{Camk2a−/−} mice (n=4 mice/group). k, l, Representative electropherogram traces and summary statistics of brain ALDH2 protein detected in different brain areas from astrocytic ALDH2 deficient mice generated by crossing Aldh2 flox with GFAPCre (Jax, 024098), Aldh2^Gfap−/− mice and their wildtype littermates, Aldh2^Gfap+/+ (n=3 mice/group). Data are presented as means ±s.e.m. Analysis was performed using unpaired two tailed student’s t test (h, j, f, l) or one-way analysis of variance (ANOVA) followed by Tukey’s test (d, e). PFC, Prefrontal cortex; CE, Cerebellum; HIPP, Hippocampus; CeA, Central nucleus of the amygdala; VTA, Ventral tegmental area; BLA, Basolateral amygdala; NAc, Nucleus accumbent.