Constructions were done using the CpG free plasmid pCpGfree-promoter (Invivogen, San Diego, CA) as the backbone to study enhancer methylation. The ANGPTL2 CpG region was amplified using forward 5’-TAAGCTCCTTCCCACGTGACCTCACAGAGTCG-3’ and reverse 5’-GATCCGACTCTGTGAGGTCACGTGGGAAGGAGCTTATGCA-3’ primers and subsequently inserted in the backbone using NsiI and BamHI restriction sites as previously described [54 , 55 (link)].
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