Extended Data Fig. 1: DNA methylation in amplicons of the target region.
a) DNA methylation at the most age-associated CpG site in PDE4C was measured by digital droplet PCR in peripheral blood, as described before (n = 80, R2 = 0.76; Han et al., BMC Biology 18,71 (2020)). b) Schematic presentation of the genomic region in PDE4C with guide RNA binding sites (blue) and the bisulfite amplicon sequencing region with corresponding methylation changes. c) Pyrosequencing measurement of HEK293T cells after epigenetic modification at PDE4C (three replicates). DNAm levels at the seven CpGs in the pyrosequencing amplicon are depicted at day 14 (***P < 0.001; **** P < 0.00001). d) Pyrosequencing of the same samples over a time course of up to 100 days confirmed that the modified DNAm is stable at this site. e) Correlation of DNAm was analyzed at the 26 neighboring CpGs within the bisulfite barcoded amplicon sequencing reads. The heatmap depicts Pearson correlations between individual CpGs. f) Heatmaps showing the complete time-course experiment upon modification with dCAS9-DNMT3A, corresponding to Fig. 1e.
