Fig. 2: The rs73613962-containing region has an enhancer signal and allele-specific enhancer activity.

a Overview of the H3K4me1, H3K27ac, and H3K4me3 chromatin modifications, the binding sites of transcription factors (Txn Factor ChIP), and DNase cluster distribution in the regions surrounding rs73613962 of the HepG2 cell line supported from the UCSC genome browser. rs73613962 is indicated by the red vertical line. b–g The dual-luciferase assays of enhancer activity for the empty vector (Control), and the plasmids with the insertion of the rs73613962-centered region (Enhancer-T and Enhancer-G). Luciferase activity is individually examined when the rs73613962-centered region is inserted in the luciferase vector in the forward (b–d) or reverse orientation (e–g). The assays are performed in two HCC cell lines, QGY-7703 (c, P < 0.0001 in Enhancer-T or Enhancer-G compared to Control, and P = 0.0026 in Enhancer-G compared to Enhancer-T; f, P = 0.0003 and P = 0.0001 in Enhancer-T or Enhancer-G compared to Control, and P = 0.0181 in Enhancer-G compared to Enhancer-T) and HepG2 (d, P = 0.0006 and P = 0.0003 in Enhancer-T or Enhancer-G compared to Control, and P = 0.0036 in Enhancer-G compared to Enhancer-T; g, P < 0.0001 in Enhancer-T or Enhancer-G compared to Control, and P = 0.0003 in Enhancer-G compared to Enhancer-T). Values are expressed as the mean ± SD, n = 3 in c, d, f, and g. ** and *** mean P-values less than 0.01 and 0.001, respectively (two-sided student’ t-test).