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Movie 1. Protein transport through the secretory pathway. COS cells expressing ts045VSVG-GFP were incubated overnight at 40°C and then shifted to 32°C to initiate ER export of the chimera. The time-lapse sequence shows movement of the VSVG-GFP from the ER to the Golgi and from the Golgi to the plasma membrane. Hirschberg et al., 1998 |
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Movie 2. Formation and trafficking of ER-toGolgi transport intermediates. COS cells expressing ts045 VSVG-GFP were incubated overnight at 40°C and then shifted to 32°C. The movie shows a 13 minute time-sequence approximately 20 minutes after temperature shift. Note that VSVG-GFP emerges from the ER at scattered sites and then moves as tubular-vesicular structures toward the juxtanuclear localized Golgi apparatus. Presley et al., 1997 |
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Movie 3. Budding of secretory cargo from the Golgi apparatus. Cells expressing ts04VSVG-GFP at 40°C were shifted to 32°C. The sequence shows the Golgi region after 50 min at 32°C. The images show numerous tubule structures budding from the Golgi that contain the chimera. Hirschberg et al., 1998 |
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Movie 4. Structure and dynamics of post-Golgi carriers. Inverted images of a ts045VSVG-GFP expressing cell obatined 50 min after shift from 40°C to 32°C. Images were captured every 2 sec. Note that VSVG buds out of the Golgi primarily in tubule transport intermediates. Hirschberg et al., 1998 |
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Movie 5. Effects of microtubule depolymerization BFA-induced Golgi absorption into the ER. HeLa cells expressing GFP-GalTase were placed on ice for 20 min and nocodazole was dded. The cells were warmed to 37°C in the presence of BFA. Very little change in Golgi morpholy occured during the first 30 minutes of BFA treatment, with no tubules observed. Beginning at 36 min and proceeding for the next 50 s, GFP-GalTase redistributed into the ER in a rapid process called 'blinkout'. These results indicate microtubule disruption delays onset of Golgi blinkout but does not affect its kenetics. Sciaky et al., 1997 |
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Movie 6. Lateral diffusion of GFP-GalTase within Golgi membranes measured by FRAP. |
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Movie 7. Rapid cycling of lipid raft markers between the plasma membrane and Golgi complex. COS cells expressing GPI-GFP were photobleached in the areas outlined and then imaged over time. Note in the top cell that the Golgi pool rapdily recovers at the expense of plasma membrane fluorescence. The bottom cell shows no recovery indcicating there was no significant new pretein synthesis during the 18 min of imaging. The data thus provides evidence for rapid constitutive cycling of GPI-GFP between cell surface and Golgi. Nichols et al., 2001 (J. Cell Biol.) |
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Movie 8. Visualization of GPI-GFP trafficking. COS cells expressing GPI-GFP were photobleached in the area surrounding the juxtanuclear Golgi region to remove plasma membrane-associated fluorescence. Subsequent time-lapse imaging revealed budding and translocation of transport intermediates carrying GPI-GFP to and from the Golgi region. Nochols et al., 2001 (J. Cell Biol.) |
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Movie 9. Golgi proteins in mitosis. A population of HeLa cells expressing GFP-GalTase were imaged over the interval shown. Note how the Golgi undergoes complete disruption during mitosis and reassembles at the end of mitosis. Zaal et al., 1999 |
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Movie 10. FLIP of GalTase-GFP expressing PTK1 cells either in mitosis or in interphase. A boxed area spanning part of both an interphase cell (left) and mitotic cell (right) was repetitively photobleached with high laser power. After each cycle of bleaching, an image of the cells was collected useing low laser power. The rapid loss of fluorescence observed throughout the mitotic cells provides evidence that during mitosis, Golgi proteins are diffusing within an interconnected membrane system with characteristics of the ER. Zaal et al., 1999 |
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Movie 11. Kinetics of membrane binding and dissociation of COPI. The Golgi region-of-interest was photobleached in cells expressing εCOP-GFP. Cells were then imaged at 5-10 sec intervals with low light illumination to monitor exchange between photobleached and non-bleached εCOP-GFP. The data revealed rapid membrane binding and release of COPI from membranes. Preslet et al., 2002 |
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Movie 12. Differential mobility of GFP-tagged lamin B receptor (a nuclear membrane protein) in ER and nuclear envelope membranes as revealed by FRAP. Ellenberg et al., 1997 |
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Movie 13. Nuclear envelope reassembly in vivo. Time-lapse sequence of COS cell expressing GFP-tagged lamin B receptor from late anaphase to cytokinesis illustrating NE reformation. Ellenberg et al., 1997 |
