Figure 5.

Verification of the cytosine methylation sites in the T. gondii genome using enzymes (HpaII/MspI) coupled with PCR. HpaII and MspI were used to digest gDNA followed by PCR or qPCR. A: At the T-IV-1336110+ site, where partial methylation was presented in tachyzoites, the methylation-sensitive enzyme HpaII did not fully cut the gDNA at this site, and PCR product appeared; however, methylation-insensitive enzyme MspI cut the gDNA at this site thoroughly, and no PCR products were obtained. B: qPCR validation was conducted after the gDNA was digested by HpaII and MspI, respectively. All results were obtained from three repetitive experiments with three replicates. T -tachyzoites, B -bradyzoites. Error bars: SEM.