Figure 8.

Some vesicular traffic continues out of cholesterol-laden organelles in NPC1 mutant cells. Images from CT60 cells transiently transfected with VAMP7-YFP were taken using streaming acquisition. VAMP7 localizes to the large abnormal organelles found in NPC, as well as VAMP7-positive, NPC1-negative vesicles that continue to move rapidly without functional NPC1. The arrow at 0.6 s points at a bright VAMP7 spot. At 4.6 s, this structure begins to extend off of the parent organelle. Between 5.5 and 5.7 s, what appears to be a fission event occurs as the tubule is severed. Bar, 1 μm. Video 9 accompanies Figure 8. Continued movement of VAMP7-positive/NPC1-negative vesicles out of cholesterol-laden organelles. Images were acquired every 0.25 s for 6 s and were played back at a rate of four images per second.