Fig 5B shows that DPN significantly increased DNA methylation at 24 and 48 h of treatment, when the MspI restriction enzyme was utilized. On the other hand, E2 significantly increased the methylation degree of CG-rich regions at 24 h of treatment, when both restriction enzymes were utilized. These data indicate that ERβ activation reverts the DNA hypomethylation status in melanoma cells and suggest that different ERβ ligands might increase the methylation of the different cytosines of the CG-rich regions (internal vs. external) in a specific way. Here, ESR2 is linked to melanoma.