Extended Data Fig. 4: Epigenetic regulators BAZ-2 and SET-6 localize at the promoter region of target genes.
From: Two conserved epigenetic regulators prevent healthy ageing
a, Co-immunoprecipitation of BAZ-2 and SET-6 using genome-edited baz-2GFP::FLAG;set-6GFP::HA worms (left) or transgenic worms expressing BAZ-2::FLAG and SET-6::HA (right). Images are representative of four independent experiments. For gel source data, see Supplementary Fig. 3. b, Representative western blots (left) and quantitative analysis (right) of H3K9 methylation levels in N2, baz-2, set-6 and baz-2;set-6 worms. Normalized H3K9 methylation levels were calculated by normalizing the ratio of H3K9 methylation and histone 3 levels to that of N2 worms. For gel source data, see Supplementary Fig. 4. c, Peaks of BAZ-2- and SET-6-binding sites in the region −1,000 bp to +1000 bp around the transcription start site (TSS). Only those peaks with a fold change of more than 2 are plotted. The y-axis indicates the average read coverage normalized to the number of uniquely mapped reads per million per genomic bin (bins = 1,000). d, Pie chart showing the distribution of overlapping BAZ-2- and SET-6-binding sites in genomic features. TTS, transcription termination site. e, f, ChIP-qPCR analysis of endogenous BAZ-2 (e) or SET-6 (f) enrichment at nuclear genes encoding mitochondrial proteins in genome-edited baz-2GFP::FLAG and set-6GFP::HA worms. ChIP-qPCR data from N2 worms were used as a control. In b, e, f, the numbers of independent experiments are shown beneath the bars; data are means ± s.e.m.; *P < 0.05; **P < 0.01; ***P < 0.001 (b, one-way ANOVA with Dunnett’s test; e, f, two-tailed t-test; see Supplementary Information for exact P values).
