Fig. 2

Haplotype-specific transcriptional activity. Luciferase reporter assays. Firefly luciferase activity was normalized to the Renilla luciferase signal and the data are shown as proportions of control vector pGL4.10-TK. a HepG2 cells with 638-bp constructs. Right panel: FOXA2 induced a haplotype-specific transcriptional activity (CGG > TAC) compared to the control pGL4.10-TK. Error bars represent the standard deviation of five experiments (n = 5) with three, three, eight, four, and four technical replicates (***p ≤ 0.005; two-tailed t-test). b HepG2 cells with 3-kb constructs. Right panel: FOXA2 induced the transcriptional activity of the inserts in a haplotype-specific way (CGG > TAC) compared to the control pGL4.10-TK. Error bars represent the standard deviation of six experiments (n = 6) with four, four, eight, eight, four, and four technical replicates (***p ≤ 0.005; two-tailed t-test). c MPHs with 3-kb constructs. Right panel: insulin reduced FOXA2-induced transcriptional activity in MPHs. Error bars represent standard deviation of four biological replicates corresponding to four livers (n = 4) with two technical replicates each (*p ≤ 0.05; **p ≤ 0.01; ***p ≤ 0.005; t-test for comparison between haplotypes within treatment; one-way ANOVA for comparisons between the treatments). a–c Left panels: in co-transfection with the pCMV6-XL5 vector without a TF insert, none of the haplotypes show transcriptional activity with respect to the control plasmid pGL4.10-TK. d HepG2 cells with 3-kb constructs. Left panel: in co-transfection with MAFK and pCMV6-XL5, none of the haplotypes show transcriptional activity. Right panel: co-transfection of FOXA2 and MAFK induced transcriptional activity from both 3-kb regions but inverted the haplotype bias compared to FOXA2 alone (TAC > CGG). Error bars represent standard deviation of six experiments (n = 6) with six, six, six, six, four, and four technical replicates (***p ≤ 0.005; two-tailed t-test)