3.6. Induction of Lipid Droplet Accumulation and Staining

HepG2 cells were seeded in a 96-well plate at a density of 8 × 10³ cells per well and incubated overnight at 37 °C. The cells were treated with 400 μM oleic acid to stimulate lipid droplet accumulation and with vehicle or the compounds at 12.5, 25, and 50 μM for 24 h. The cells were fixed with 4% paraformaldehyde for 30 min and then stained with 1 µg/mL Nile red and 0.1 µg/mL DAPI for 15 min. The fluorescence of each sample was measured using a 550 Bio-Rad plate-reader (Bio-Rad) [49].