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Fig. 3. ERGIC stationary structures in the periphery and in the Golgi area are equivalent. (A) Time points from supplementary material Movie 5 showing the relocalization of GFP-ERGIC-53 to the ER when cells are treated with H89. The arrow points to a peripheral ERGIC structure that loses fluorescence and disappears without moving to the Golgi area. (B) Time series from supplementary material Movie 6 showing the recovery of the same cell after H89 washout. The arrow points to a peripheral ERGIC structure that gets brighter during recovery but does not move to the Golgi area (arrowhead) which is refilled at the same time as peripheral ERGIC structures appear. (C) NEM blocks H89-induced recycling of GFP-ERGIC-53 to the ER. HeLa cells stably expressing GFP-ERGIC-53 were treated with H89 for 20 minutes in the presence or absence of NEM or left untreated (control), fixed with paraformaldehyde, stained for Sec13 (red), and analyzed by confocal microscopy. Insets show higher magnifications. Bar, 5 µm. Time is indicated as minutes.seconds.
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