Fig 1: Levels of AcH3, ALDH1A1, and RA-RARa expression in the PFC of VPA-exposed offspring following MS-275 treatment. (A) Western blot and quantification analyses of AcH3 and total H3 protein in the nuclear fraction of PFC from the CON + Saline, VPA + Saline, and VPA + MS-275 groups, as normalized to PCNA; the level of histone acetylation was measured as the ratio of AcH3 to H3 (n = 3 per group). (B) ALDH1A1 mRNA expression in the PFC from the CON + Saline, VPA + Saline, and VPA + MS-275 groups, as detected using qPCR and normalized to GAPDH (n = 10 per group). (C) Western blot and quantification analyses of ALDH1A1 protein in the PFC of offspring from the CON + Saline, VPA + Saline, and VPA + MS-275 groups (n = 3 per group). (D) ALDH enzyme activity in PFC tissue from the CON + Saline, VPA + Saline, and VPA + MS-275 groups was determined using the ALDH Assay Kit (n = 10 per group). (E) RA levels in the PFC from the CON + Saline, VPA + Saline, and VPA + MS-275 groups, as quantitated using UPLC-MS/MS (n = 5 per group). (F) Western blot and quantification analyses of RARa protein in the PFC from the CON + Saline, VPA + Saline, and VPA + MS-275 groups (n = 3 per group). (G) ChIP-qPCR analyses of the enrichment of AcH3 on the promoter region of the ALDH1A1 gene in the PFC from the CON + Saline, VPA + Saline, and VPA + MS-275 groups (n = 3 per group). Each experiment was repeated at least three times. The values are the means ± SEMs (A,C–G) or median (P25–P75) (B). One-way ANOVA with Bonferroni post hoc test (A,C–G), Kruskal–Wallis H test with Nemenyi post hoc test (B), *P < 0.05, **P < 0.01, ns, not significant; VPA, valproic acid; PFC: prefrontal cortex.