Fig. 2: Reporter assay of the HAL-promoter region containing putative CEBP- and Forkhead-binding elements.

a Sequences of the wild type and mutant HAL-promoter region used for the reporter assay. The 5’-flanking region of HAL (between −90 and −44 bp) contains two putative CEBP-binding elements (CBE-1 and CBE-2) and a putative Forkhead-binding element (FBE). Substitutions in the binding elements are underlined and shown in bold. b Reporter activity of the wild-type and mutant HAL-reporter plasmids in response to β-catenin-9 siRNA. The activity of each reporter plasmid in the cells treated with β-catenin siRNA was divided by that with control siRNA. Dual reporter assay was carried out using pRL-null plasmid for the normalization of transfection. c The effect of CEBPA, FOXA1, and FOXA3 over-expression on the reporter activity. HepG2 cells were transfected with wild type HAL-reporter plasmid and the indicated transcription factors. The y-axis represents relative reporter activity compared to the mock reporter plasmid. d The effect of siRNA for CEBPA, FOXA1, and FOXA3 on the reporter activity. Hep3B cells transfected with the wild type reporter plasmid were treated with the indicated siRNAs. The y-axis represents the relative reporter activity compared to the mock reporter plasmid. Unless specified otherwise, data are represented as the mean ± SD of three independent cultures. Statistical significance was determined by Dunnett’s test. *p < 0.05, **p < 0.01 vs Empty or siCtrl.