Extended Data Fig. 4: Further analysis of ATAC-seq and 4C-sequencing data.

a) Violin-plot of ATAC-seq signals comparing chromatin accessibility at all CpGs and hypermethylated bystander positions of FHL2 (delta DNAm >10%; n = 2, P < 10⁻¹⁵, unpaired t-test), in analogy to Fig. 3a. b) Multivariate kernel density estimate of ATAC-seq and DNA methylation changes (dCAS9-DNMT3A). While differentially methylated positions exhibited in tendency higher chromatin accessibility the overall correlation was rather low. In analogy, we analyzed the 4389 age-hypo and 5328 age-hypermethylated CpGs. c) Cis-interactions (on the same chromosome) of 4C-Sequencing with around 20 million reads per sample. Particularly the two viewpoints at positively age-correlated regions in PDE4C and FHL2 exhibited cis-interactions spanning a mean of 271 and 672 kbp, respectively. In contrast, the 4 C reads of age-hypomethylated region in MEIS1-AS3 comprised only 89 kbp. d) Trans-interactions (across different chromosomes) of 4C-sequencing. The viewpoints at positively age-correlated regions like PDE4C and FHL2 exhibited trans-interactions spanning a mean of 6.5 and 13.0 mbp, respectively. e) Bystander effects of FHL2 (abs. difference beta-value > 0.1; n = 2) become enriched with increasing coverage in 4C-sequencing of FHL2 interacting sites (n = 3, different symbols correspond to replica; R² = 0.81, linear model P < 10⁻¹⁵).