Figure 4

Validation of MeDIP-Seq data by bisulfite sequencing. A total of 5 genes were selected randomly to validate MeDIP-Seq data by bisulfite PCR in blactocyst-stage cloned and IVF embryos. CpG islands of upstream 2 k of all selected genes were predicted by MethPrimer. No of circles in each horizontal line represent no of CpGs present in a particular regions. Unfilled (white) and filled (black) circles represent unmethylated and methylated CpGs, respectively. Horizontal lines of circles represent one separate replicate that was sequenced. Lollipop diagrams were generated by BIQ Analyzer software. For each sample, the methylation data were analyzed by computing the percentage of methylated CpGs of the total number of CpGs. (A) DNA methylation status of upstream 2 kb of PEG10. F2-R2 region was chosen as the analysed region. 16 circles in each horizontal line represent 16 CpGs of PEG10. (B) DNA methylation status of upstream 2 kb of IGF2R. F2-R2 region was chosen as the analyzed region. 13 circles in each horizontal line represent 13 CpGs of IGF2 (C) DNA methylation status of upstream 2 kb of MDM2. F1-R1 regions was chosen as the analyzed region. 11 circles in each horizontal line represent 11 CpGs of MDM2. (D) DNA methylation status of upstream 2 kb of TCF7. F1-R1 region was chosen as the analyzed region. 20 circles in each horizontal line represent 20 CpGs of TCF7. (E) DNA methylation status of upstream 2 kb of COL4A1. F2-R2 region was chosen as the analyzed region. 15 circles in each horizontal line represent 15 CpGs of COL4A1.