Lipid Droplet Quantification in Huh7 Cells

Lipid droplet assay was performed according to a previous method using a BSA-conjugated oleic acid system in Huh7 cells as described previously [22 (link)]. Briefly, cells seeded in μClear® 96-wells plates (Greiner Bio-ONE, Frickenhausen, Germany) were treated with oleic acid and the tested sample or DMSO for 18 h. Paraformaldehyde was used to fix the cells, which were stained with 2 μg/mL Hoechst 33342 and 1 μg/mL BODIPY® 493/503. High Content Imaging (HCS) instrument was used to take and analyze images of the nuclei and lipid droplets (ImageXpress Micro System, Molecular Devices, Sunnyvale, CA, USA). The diameter settings were 8–25 μm for the nuclei and 0.5–2 μm for the lipid droplets.