Figure 9. ORP4 expression had no effects on cholesterol production.

A. Huh7.5 cells were transfected with the indicated amount of ORP4 expression plasmid. Forty-eight hours post transfection, cell culture supernatants were collected, while the cells were harvested for cell lysates. Cell culture supernatants and cell lysates were used to determine secreted cholesterol (open bar), the total intracellular cholesterol (closed bar), respectively. Standards were used to calculate the absolute concentration of cholesterol in both samples. The data are Means ± SD of triplicates and are representative of at least three independent experiments. B. Similar transfections were performed, the cell lysates were analyzed for SREBP1c and ORP4 expression by Western blotting. β-actin was included as an equal loading control.