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Fig. 2
Sma and Mad homologs (SMADs)–forkhead box protein H1 (FOXH1) complex–controlled HNF4α regulates expression of coagulation factors in LPCs. (A) Chromatin immunoprecipitation (ChIP) assay was performed to measure HNF4α binding to the promoter of coagulation factor 2 (F2) and 5 (F5) genes in HepaRG and bipotential murine oval liver (BMOL) cells. (B) Quantitative PCR and western blotting were used to measure mRNA expression of coagulation factors in HepaRG and BMOL cells with or without HNF4α RNA interference (RNAi). (C) ChIP assay was performed to measure SMAD2/3, SMAD4, and FOXH1 binding to the promoter of hnf4α genes in HepaRG and BMOL cells. (D) Co-immunoprecipitation (Co-IP) was performed to measure SMADs binding to FOXH1 in HepaRG cells. The expression of the cyclophilin A (PPIA, peptidylprolyl isomerase A) was used as loading control in ChIP assay. Tubulin was used as loading control in western blotting. *p < 0.05, **p < 0.01, and ***p < 0.001
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