Figure 4.
Methylation status of the DMRs in blastocysts lacking both maternal and zygotic Dnmt1. (A) A schematic representation of the flow of the experiment. [Dnmt12lox/2lox, Zp3-Cre] females were crossed with Dnmt1c/+ males, and then the obtained E3.5 blastocysts were genotyped by PCR with primers that specifically amplify the Dnmt1c allele. DNA from blastocysts of the same genotype (74 blastocysts of Dnmt11lox/c and 111 blastocysts of Dnmt11lox/+) was pooled and subjected to bisulfite sequencing. (B) Methylation status of the H19, Rasgrf1, Peg3, and Snrpn DMRs in wild-type (top) and mutant blastocysts (middle and bottom). (Middle) Blastocysts lacking maternal Dnmt1 (Dnmt11lox/+) showed a partial reduction in methylation at the normally methylated allele of all DMRs. (Bottom) Blastocysts lacking both maternal and zygotic Dnmt1 (Dnmt11lox/c) showed near complete loss of methylation at the normally methylated DMR alleles. (JF1) JF1-derived allele; (dom) domesticus-derived allele.