Fig. 2.

Cholesterol-dependent internalization of NPC1L1-GFP^turbo is ameliorated by GM3S depletion. A: Control HEK293T and GM3S KO cells were seeded in 0.001% poly-l-lysine-coated 35-mm glass-bottom dishes on day 0 and transfected with NPC1L1-GFP^turbo on day 1. The medium was replaced with medium containing 5% lipoprotein-deficient serum and 2 µM compactin on day 2 to deplete cellular cholesterol, and cells were supplemented with 60 µg/ml cholesterol with or without ezetimibe treatment on day 3. Time-lapse images were taken by confocal microscopy. Representative images are shown. B: Quantification of plasma membrane-localized NPC1L1-GFP^turbo in the cells shown in panel A. Intensity at time zero was defined as 100%. *P < 0.05 for comparison of control versus GM3S KO.