Huh7 GNMT Promoter-Luciferase Assay

Huh7 GNMT promoter-luciferase (H7GPL) cells^{75 (link)} were seeded at 96-well plates, 5 × 10³ cells in each well. H7GPL cells were treated with 2.5, 5, 25, 50, 100, 250, 500 μM AAI or 0.1% DMSO (vehicle negative control) in duplicate for 16 hrs. Treated H7GPL cells were lysed for luciferase activity measurement by using the One-Glo^TM Luciferase Assay System (Promega, E6120) and detected in a luminometer (BioTek Ins., Synergy HT). The reporter luciferase activity were normalized by cell density measured by adding alamarBlue® reagent (Thermo Scientific/Pierce Biotechnology., Rockford, lL, USA). Values were presented as the relative luciferase activity, fold increase over vehicle control.