Figure 2.

YY1 KD effects during spermatogenesis. Sperm DNA was isolated individually from the F1 mice (pSico-YY1, Pro-Cre) and the littermate controls (+, Pro-Cre). The individual F1 mice are indicated by different numbers, while the control littermates are indicated by numbers followed by C (1C, 5C, 8C and 14C). (A) Individual DNA samples from the 3-month-old set were treated with the bisulfite conversion method, followed by the restriction-enzyme-based method—COBRA. Digestion and lack of digestion by TaqI and ClaI indicate the methylated (M) and unmethylated (U) status, respectively, of a given CpG site in the original DNA as shown in the digestion of Peg3, Xist, Tsix and H19. In contrast, this is opposite for the HphI digestion, which recognizes TpG as shown in the digestion of Nespas. Digestion by this enzyme indicates the unmethylated status of a CpG site, whereas lack of digestion indicates methylation. (B) A subset of the 6-month-old mice is also shown in the same manner as the above. The results for H19 (8#C and 9) are not available due to the unsuccessful PCR amplification.