Hnf4a Promoter Transactivation Assay

The mouse Hnf4a promoter sequences were inserted into the NheI/HindIII site of the luciferase pGL3-Basic vector (Promega, USA). The primary hepatocytes were divided into three groups: the control group (transfected with reporter vector pGL3-Hnf4a promoter and control plasmid), Hnf4α group (transfected with reporter vector pGL3-Hnf4a promoter and Hnf4a plasmid), and Hnf4α + lauric acid group (transfected with reporter vector pGL3-Hnf4a promoter and Hnf4a plasmid, then stimulated with lauric acid). All groups were cotransfected with target vectors and pRL-TK (Promega, USA). After 24 h of incubation, the cells were treated with DMSO or lauric acid (0.5 mmol/L) for 24 h. Next, the dual-luciferase reporter assay was performed according to the manufacturer’s instructions (Promega, USA).