The mitochondrial adaptation in AT-1 Tg mice is driven by a specific epigenetic activation. (A) Mass spectrometry quantification of posttranslational modifications of histone proteins in the hippocampus of AT-1 Tg mice. Global changes on H3K27/K36 are shown on the left and changes in the acetylation and methylation profile of H3K27 are shown on the right (WT, n = 3; AT-1 Tg, n = 3). (B) Western blot assessment showing acetylation and methylation of H3K27 in AT-1–overexpressing H4 cells. Selected images are shown on the left panel and quantification (n = 4) is shown on the right. (C) ChIP analysis of SLC33A1, ACLY, and ACSS2 after immunoprecipitation (IP) with an anti-H3K27ac antibody. IP was done in H4 cells. (left) Sheared DNA; (right) ChIP-amplification. Analysis was performed in quadruplicate. (D) ChIP-qPCR showing H3K27 acetylation of SLC25A1 and ACLY in H4 cells. Results are expressed as fold of H4 (-) (H4(-), n = 5; H4(AT-1WT), n = 5). (E) ChIP-qPCR showing H3K27 acetylation of Slc25a1 and Acly in total hippocampal tissue. Results are expressed as fold of WT (WT, n = 3; AT-1 Tg, n = 3). (F) ChIP-qPCR showing H3K27 acetylation of Slc25a1 and Acly in isolated adult neurons. Results are expressed as fold of WT (WT, n = 3; AT-1 Tg, n = 3). IP in D–F was performed with an anti-H3K27ac antibody. *, P < 0.05; **, P < 0.005; #, P < 0.0005. Student’s t test and one-way ANOVA followed by Tukey-Kramer multiple comparisons test. Bars represent mean ± SD.
