Extended Data Figure 7: Characterization of the regulatory CpG island CpG43.

a, Snapshot of Genome Browser view of genomic DNA around the mir-200ba429 cluster taken from NCBI37/mm9. Tet2 ChIP was obtained from GSE41720, sample GSM1023124. Shaded rectangles indicate mir-200ba429 and CpG43. b, ChIP–PCR of the indicated histone marks in a region adjacent CpG43. Data were obtained from three independent cultures and are presented as average ± s.d. P values from unpaired t-tests are indicated. c, Expression levels of H3 histone marks in cells of the indicated genotypes measured by western blot with H3 and calnexin as loading controls. d, 3C data for the genomic region adjacent to CpG43 analysed in Fh1^fl/fl cells. The positions of CpG30 and CpG43 and of the predicted restriction sites are indicated. Results were generated from two independent cultures. e, DNA methylation of CpG43 assessed by qPCR using OneStep qMethyl kit. Data were obtained from three independent experiments and normalized to methylation levels of the region in Fh1^fl/fl cells. Data are presented as average ± s.e.m. f, ChIP–PCR of Tet binding to CpG43. Data were obtained from three replicates and are presented as average ± s.d. g, 5hmC nuclear staining assessed by immunofluorescence using 5hmC antibody. Nuclear staining was quantified using Image J and an average of 120 cells was used per genotype. P values from one-way ANOVA. Representative images of 5hmC staining are shown. DAPI is used to stain nuclei. Scale bar, 20 μm. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001. Western blot source images are presented in Supplementary Fig. 1. Source data are presented in Supplementary Table 2.