Luciferase Assay for PPARα Activity

For luciferase assays, HepG2 cells were seeded at a density of 1 × 10⁴ cells in a 96-well plate. Cells were transfected with lipofectamine transfection reagent (Thermo Fisher Scientific, Rockford, IL, USA), and plasmids were used for transfection with the PPRE-X3-TK-LUC plasmid (Dr. Christoper K. Glass, University of California, San Diego, CA, USA) and PPARα expression vectors (Dr. Han Geuk Seo, Konkuk University, Seoul, South Korea). After transfection for 24 h, cells were treated with WY14643 (a PPARα agonist) [71 (link)] or SSC for 6 h. Luciferase activity was measured using the One-Glo Luciferase Reporter Assay System (Promega, Madison, WI, USA) and a luminescence plate reader (Berthold Technologies GmbH & Co., Bad Wildbad, Germany.