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. 2003 Aug;23(16):5594–5605. doi: 10.1128/MCB.23.16.5594-5605.2003

FIG. 3.

FIG. 3.

Expression of Dnmt3a/3b proteins in 7aabb cells restores DNA methylation. (A to D) Methylation of repetitive sequences. Genomic DNA from the indicated ES cell lines was digested with HpaII (A to C) or MaeII (D) and hybridized to the indicated probes. DNA from J1 cells digested with MspI was used as a control for complete digestion. (E) Analysis of the methylation status of the major satellite repeating unit by bisulfite sequencing. Genomic DNA from J1 and 7aabb cells, as well as from stable cell lines expressing Dnmt3a, Dnmt3a2, Dnmt3b1, and Dnmt3b3, was analyzed. The methylation status of six CpG sites from 8 to 12 individual clones is shown schematically (black circles represent methylated sites), and the percentages of methylated CpG sites are indicated in parenthesis. (F to I) Methylation of unique genes. The genomic DNA samples described in panels A to D were digested with BamHI and HhaI (F and H), EcoRI and HpaII (G), or EcoRV and HhaI (I) and hybridized to probes corresponding to the 3′ region of β-globin (F), the 5′ region of Pgk-1 (G), an exon of Pgk-2 (H), or the 5′ region of Xist (I). DNA from J1 cells digested with BamHI alone (F and H) or EcoRI alone (G) was used as controls.