Figure 4
From: Zebularine regulates early stages of mESC differentiation: effect on cardiac commitment
Analysis of gene promoter’s methylation under zebularine treatment. (a) MSP of pluripotency and cardiac lineage markers showing that the promoters of both Oct3/4 and Nanog shifted from unmethylated (U) to methylated (M) state; the promoter of Nkx2.5 becomes unmethylated after zebularine treatment and no promoter methylation was detected in any of the other cardiac markers analyzed. (b) Bisulfite sequencing from five clones of Gata4 CpG island for Control, zebularine and AzadC-treated EBs, indicating that the promoter remains unmethylated in 15 CpG sites. (c) Bisulfite sequencing from five clones of Nkx2.5 promoter revealed that zebularine unmethylated almost all of the 43 CpG sites analyzed. The percentage of methylation is presented as the percentage of filled circles over total circles for a given CpG site from each clone analyzed: 
=0%; 
=40%; 
=60%; =80%; 
=100%
