Transient transfection reporter assays were all performed in 96-well format. Transfection complexes were formed by incubating 100 ng of each individual promoter construct with 0.3 µL of Fugene 6 transfection reagent and Opti-MEM media in a total volume of 5 µL and incubated for 30 min. Transfection complexes (5 uL) were added to 10,000 HepG2 cells in 96-well format that had been seeded 24 h prior to transfection in a white tissue-culture treated plate.
