Figure 4

Methylations of PITX2, TLX3 , OTX2 and MNX1 genes validated using the combined bisulfite restriction analysis. Bisulfite converted genomic DNA was amplified with gene-specific primers followed by digestion with a methylation sensitive enzyme BstUI (CG↓CG). Methylation status was measured in placental tissue (P), maternal peripheral blood (M) and chorionic villus (C). The expected enzyme-digested pattern of the PCR products in which CpG islands were fully methylated was depicted on right side.