Fig. 9. Accumulation of cholesterol in LGP85-induced large vacuolar compartments. COS cell cultured in DMEM supplemented with 10% FBS were transiently transfected with LGP85 and fixed after 36 hours after transfection. Cells were immunostained for LGP85 (B) and cytochemically stained with filipin (C) to detect cholesterol. To examine further whether cholesterol accumulated in the LGP85-induced large swollen vacuoles is derived from one containing into cellular membranes or into LDL, COS cells cultured in DMEM supplemented with 10% LPDS instead of 10% FBS for 2 days were transiently transfected with LGP85, fixed at 36 hours after transfection, and stained for LGP85 (F) and cholesterol (G). The right columns (D,H) show the merged images of LGP85 (red) and cholesterol (blue), and the left columns (A,E) show the corresponding phase-contrast micrographs. Bars, 20 µm.