Figure 4.

Effect of MXC on CpG sites in PAPP-A gene in adult ovary. Bisulfite sequencing was conducted to examine the regions −566 to −329 and +8 to +300 upstream and downstream, respectively, from the transcription start site of the PAPP-A gene (shown above are data from the +8 to +300 group). Eight separate clones were analyzed from two or three animals from each treatment group. All predicted CpGs that were converted to thymine are shown as open circles and methylated cytosines that remain as cytosines are shown as filled circles. Although no changes were observed in the upstream region of the start site, numerous alterations were observed in the 16 CpGs in the +8 to +300 region. Only one predicted CpG site was methylated in control ovaries. However, numerous CpGs were methylated in ovaries exposed to 20 μg/kg·d and 100 mg/kg·d MXC.