Figure 3.

Effects of MXC on the global DNA methylation in ovaries at PND 50–60. Methylation-sensitive AP-PCR was used. Four micrograms of ovarian genomic DNA were digested with RsaI (R) or RsaI with methylation-sensitive HpaII (H) or -insensitive MspI (M) restriction enzymes, followed by PCR with 10 degenerate primer sets designed to amplify methylation sites (five representative PCR product sets are shown, 1–5; each panel shows one set). C, Control; MXC, 100 mg/kg·d MXC. See Materials and Methods for details. The H-digested PCR products that were differentially amplified between control and MXC (arrowheads; enlarged inset, and arrow) were identified and listed as potential candidates for further investigation (Table 1); (n = 3 animals per treatment). All experiments were repeated three times.