Functional assays for g.3131C > T SNP (rs17248720). A) EMSA assay carried out with probes containing the C or T allele for the g.3131 SNP (rs17248720) in the LDLR gene. NE, no HepG2 extract added. E, nuclear extract from HepG2 was added, ×10, ×20, ×50 and ×100 mean 10, 20, 50 and 100 times excess of unlabelled C-allele oligonucleotide. B) The inverse of band densities from the EMSA shown in Figure 1A were plotted against the excess of unlabelled allele C oligonucleotide. C) Influence of g.3131C > T (rs17248720) variant on LDLR promoter strength in transient transfection assay in FBS supplemented media. Fragments containing, none, either C or T allele were cloned in the promoterless plasmid pGL3-Basic and transfected to FBS supplemented media cultures of HepG2. Measurements of luciferase activity were assayed in cell extracts 36 h after transfection. The results were expressed as the media of three experiments carried out in triplicate considering pGL3-Basic expression as value 1.