The region directly surrounding HERC2 rs12913832 acts as a melanocyte-specific enhancer. (A) Formaldehyde assisted identification of regulatory elements (FAIRE) demonstrates low nucleosome occupancy at the rs12913832 region in HEMn-LP and HEMn-DP cells. (B) ChIP-qPCR of acetylated histone H3 demonstrates active chromatin marks at the rs12913832 region in HEMn-LP and HEMn-DP cells. (C) ChIP-qPCR of histone H3 mono methylated on lysine 4 demonstrates that the rs12913832 region in HEMn-LP and HEMn-DP cells has enhancer potential. (D) ChIP-qPCR of histone H3 acetylated on lysine 27 demonstrates that the enhancer at the rs12913832 region in HEMn-LP and HEMn-DP cells is active. The enrichments displayed are relative to NDN. ChIP values for histone H3 marks are normalized to histone H3 occupancy. (E) Luciferase reporter assay demonstrates differential melanocyte enhancer activity for the rs12913832 region. The rs12913832 region from HEMn-LP (C-allele) and HEMn-DP (T-allele) was inserted into a luciferase reporter plasmid and transfected into HEK293 or G361 melanoma cells. Luciferase expression is normalized to Renilla luciferase expression. Data are represented as mean ± SEM; (*) p < 0.05; (***) p < 0.005.
