Fig. 7: Pontin and Tip60 complex is important for autophagic occurrence through the regulation of FOXO3a target genes.
From: Pontin arginine methylation by CARM1 is crucial for epigenetic regulation of autophagy
a Autophagic flux was analyzed in MEFs after knocking down with shRNA for Tip60 (shTip60) or non-specific shRNA (shNS) in the presence or absence of Bafilomycin A1 (200 nM; 2 h) under glucose starvation. The LC3-II/β-actin ratio is indicated. b Quantitative RT-PCR analysis of methylated Pontin-dependent genes and independent genes in MEFs by knocking down with either shNS or shTip60 in the absence or presence of glucose starvation. Bars, mean ± s.e.m; n = 3; **p < 0.01, ***p < 0.001, NS non-significant. Statistics by two-tailed t-test. c ChIP assays were performed using anti-FOXO3a, anti-Pontin-me, anti-H4ac, anti-Tip60, and anti-IgG antibodies on FOXO REs in MEFs by knocking down with either shNS or shTip60 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. d Quantitative RT-PCR analysis of methylated Pontin-dependent autophagy and lysosomal genes in Pontin WT or RA MEFs with or without Tip60 knockdown. Bars, mean ± s.e.m; n = 3; NS non-significant. One-way ANOVA with post hoc Tukey’s test. e Immunoblot analysis of LC3 levels in Pontin WT or RA MEFs with or without Tip60 knockdown. The LC3-II/β-actin ratio is indicated. f Schematics for the transcriptional activation of FOXO3a and methylated Pontin-dependent autophagy genes regulated by Tip60 with increased H4 acetylation upon glucose starvation. Glucose starvation-induced methylated Pontin by CARM1 at R333 and R339 residues binds FOXO3a through a methyl binding domain of FOXO3a along with Tip60 following increased H4 acetylation. Source data are provided as a Source Data file.
