Figure 4.

MODY1/HNF4A Mutation Results in Loss of Ability to Activate Downstream Target Promoters in Hepatic and Pancreatic β Cells

(A–D) Luciferase assays were performed to evaluate effects of WT or p.Ile271fs (Mut) HNF4A on the (A) HNF1A promoter, (B) APOB promoter, (C) AFP enhancer activity in hiPSC-derived hepatic cells, or (D) HepG2 cells. For (A–C), data are represented as mean ± SD n = 3, representative of two independent experiments. For (D), data are represented as mean ± SD of n = 12 from three independent experiments. *p < 0.05 versus GFP control in all hiPSC lines by two-way ANOVA; ^#p < 0.05 versus GFP control in mutant hiPSC lines only by two-way ANOVA. **p = 0.01 versus GFP control by Student's t test.

(E) Chromatin immunoprecipitation qPCR analysis of HNF4A binding onto HNF1A promoter in EndoC-βH1 cells.

(F) Small interfering RNA-mediated knockdown of HNF4A in EndoC-βH1 cells.

(G) Luciferase assay evaluating HNF1A promoter activity upon knockdown of HNF4A and rescue. For (E–G), data are represented as mean ± SD of n = 12 from three to four independent experiments. *p < 0.05 versus IgG/GFP control as indicated by Student's t test.

See also Figure S5.