Fig. 4
From: Hit-and-run epigenetic editing prevents senescence entry in primary breast cells from healthy donors
DNA methylation is consistently maintained at HIC1, RASSF1 and CDKN2A. a Scatter plot of DNA methylation from all EPIC array probes comparing donor 1 and donor 2 early passage primary myoepithelial cells (n = 1). The probes from the target genes highlighted on the plot are those that were found to be significantly hypermethylated at day 10 after dCas9 3A3L vs. 3A3L∆ targeting as shown in Fig. 2e. b Scatter plot of DNA methylation from all EPIC array probes comparing donor 1 and 2 primary myoepithelial cells 38 and 37 days respectively after targeting dCas9 3A3L to HIC1, PTEN, RASSF1 and CDKN2A. The same probes from a are highlighted (n = 1). c–f Targeted bisulfite sequencing at c HIC1, d PTEN, e RASSF1 and f CDKN2A. The localisation of the bisulfite amplicon (dark grey bar) and gRNA (grey arrows pointing towards the PAM sequence) in relation to CGI (green bar) and TSS of genes is shown. Each rectangle represents the methylation % indicated by the colour key of a single CpG and each line is data from a single replicate. The top 5 lines show data from early passage (passage 2) primary myoepithelial cells from donors 1, 2 and 3, the bottom 7 lines show data from primary cells 37–41 days after transfection with dCas9 3A3L targeting HIC1, RASSF1, PTEN and CDKN2A. Top lines of data from donor 1 early passage and dCas9 3A3L targeted is from the same samples as the EPIC array data in Fig. 4a, b. Bisulfite sequencing data from donor 2 is from the same samples as the EPIC array data shown in Fig. 4a, b
