FIGURE 1.

The transport of NPC1L1 to the PM requires microfilament integrity and dynamics. A, CRL-1601/NPC1L1-EGFP cells were pretreated with the indicated drugs for 30 min to disturb the cytoskeleton. These compounds were present throughout all time durations. The cells were then incubated in cholesterol-depleting medium to reduce cellular cholesterol for various time durations. At the indicated time points, the cells were fixed and examined by confocal microscopy. Images show the cells representative of the whole cell population at each indicated time point. Ctr, control; Cyt D, cytochalasin D; Lat B, latrunculin B; Jasp, jasplakinolide; Noco, nocodazle; Colc, colchicine. Bar, 10 μm. B, quantification of the intracellular localization of NPC1L1-EGFP shown in A. Error bars represent standard deviations. C, CRL-1601/NPC1L1-EGFP cells were treated as shown in A. After depletion of cholesterol for 30 min, the cells were collected and subjected to a surface biotinylation assay. Immunoblot analysis was carried out using the indicated antibodies. Results shown are representative of three independent experiments.