Fig. 3.

RNR inhibition increases 5-aza-2′-dC-mediated genome-wide DNA demethylation in MEFs. a Bar chart displaying average genome-wide CpG methylation levels for the indicated 72 h treatments filtered for CpGs with at least 5X sequencing coverage by RRBS across all samples. Label color reflects the various treatment groups. Subscripts (A and B) indicate replicates where applicable. Treatment concentrations are: LowAza (5-aza-2′-dC 0.2 μM), HighAza (5-aza-2′-dC 10.0 μM), and Resv (resveratrol 20 μM). b Heat map of unsupervised hierarchical clustering of DNA methylation levels for all autosomal CpGs assayed by RRBS in MEFs treated with the indicated chemicals for 72 h as in a with at least 5X sequencing coverage across all samples. A methylation level of 1 indicates 100 % methylation, while 0 represents complete absence of methylation. c Heat maps as in b but for subsets of autosomal CpG sites partitioned into four groups representing different DNA methylation levels in the untreated control samples; (i) 0.75–1.0, (ii) 0.50–0.75, (iii) 0.25–0.50, and (iv) 0–0.25. In each case, the combination of RNR inhibition with 0.2 μM 5-aza-2′-dC clusters away from all other samples, but together with the high dose of 5-aza-2′-dC, and is more demethylated. d Histograms display DNA methylation distributions for all autosomal CpGs as in b for indicated treatments and replicates. Data for additional replicates can be found in Additional file 8: Figure S7C/D