Filipin Staining Assay

To measure for cholesterol, HepG2 cells were cultured, seeded, and treated as described in Cell Culture and Treatment section. After 18 h of further incubation at day 2, cells were washed with DPBS 3 times, and fixed with 3.7% paraformaldehyde for 1 h. Following washing with DPBS 3 times, glycine solution dissolved in DPBS (100 μl) was added to each well to remove residual paraformaldehyde. Filipin solution (50 μg/ml) was then added to each well and incubated for 2 h at room temperature, followed by rinsing with DPBS 3 times. Fluorescence intensity was measured at 360/480 nm (excitation/emission) using a SpectraMax M2 microplate reader (Molecular Devices).