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. 2020 Dec 8;11:6297. doi: 10.1038/s41467-020-20080-9

Fig. 6. Methylated Pontin and Tip60 activate FOXO3a target genes with histone H4 acetylation.

Fig. 6

a Effect of Pontin WT or RK on 6x DBE (FOXO RE)-luciferase reporter. Bars, mean ± s.e.m; n = 3; ***p < 0.001. Statistics by two-tailed t-test. b Effect of FOXO3a WT, or F640L/F642L mutant on 6× DBE-luciferase reporter. Bars, mean ± s.e.m; n = 3; ***p < 0.001, NS non-significant. Statistics by one-way ANOVA with post hoc Tukey’s test. c Heatmap showing methylated Pontin near the TSS (above) or distally located to TSS (below). The data on H3K4me1, H3K4me3, and H3K27ac were obtained from MEFs under normal conditions. d Profiles of methylated Pontin (Normal), methylated Pontin (Glc starv.), H3K4me1, H3K4me3, and H3K27Ac ChIP-seq occupancy in the locus of the Map1lc3b gene. Gray boxes indicate possible Pontin binding sites. e ChIP assays were performed using anti-FOXO3a, anti-Pontin-me, anti-H4ac, anti-Tip60, and anti-IgG antibodies on FOXO REs in WT or Carm1−/− MEFs in the absence or presence of glucose starvation. Bars, mean ± s.e.m.; n = 3; **p < 0.01, ***p < 0.001. Statistics by two-tailed t-test. f Co-immunoprecipitation assay between FOXO3a and Tip60 in the absence or presence of Pontin. g Co-immunoprecipitation assay between Tip60 and Pontin WT or RK mutants in the absence or presence of glucose starvation. Source data are provided as a Source Data file.