Fig. 4

Follistatin treatment triggers changes in DNA methylation within intron 1 of CDX2 gene in SCNT blastocysts. A CDX2 gene map showing the genomic position of fragment I (+3030 to +3710). B Individual CpG methylation profile in fragment I, genomic DNA, from untreated control and follistatin treated groups (n= 6 replicates, n= 15 blastocysts/group), was subjected to bisulfite treatment followed by PCR amplification, cloning, transformation and finally colony PCR to select positive clones (n= 8 clones/replicate) for Sanger sequencing. In the methylation grid, rows represent the sequenced clones (Clones are arranged according to their replicates, replicate number are shown in the left) and columns represent CpG sites. Unmethylated CpG sites are represented by open circles while methylated CpG sites are represented by closed circle. C Individual CpG sites with ≥ 20% methylation, at least in one group. Values show the methylation percentage of each CpG site, and those values marked with an asterisk are significantly different (*P < 0.05)