Lipid droplet breakdown requires Dynamin 2 for vesiculation of autolysosomal tubules in hepatocytes

Video 3.
Live-cell microscopy of a Hep3B hepatocyte expressing LAMP1-mCherry and starved for 2 h in HBSS + Ca²⁺/Mg²⁺ + 10 mM Hepes, followed by 30 min treatment with 40 µM Dynasore and subsequent drug washout and replacement with media containing 10% FBS. Notice the marked reduction in overall number and size of LAMP1-positive tubules over time after Dynasore washout. Images were acquired every 20 s using epifluorescence microscopy (Axio Observer; Carl Zeiss). Movie is displayed at 10 fps and represents time elapsed between 10 and 45 min after drug washout. Contrast was enhanced by histogram normalization in ImageJ (National Institutes of Health) for better visualization of the LAMP1-positive tubules.