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Figure 8


Fig. 8. Colocalization of secretory organelle markers with dilated ER exit site (ERES) membranes. (A) COS7 cells, cotransfected with CFP- or YFP-labeled VSVG (red) and Srβ-YFP, Sec13-YFP, Sec31-CFP, p58/ERGIC53-YFP and GRASP65-DiHcRED (green) were shifted to the permissive temperature (34°C) after overnight incubation at 39.5°C, followed by 15 minutes at 4°C when nocodazole and brefeldin A (BFA) were added. Confocal images were captured 30 minutes after the temperature shift. (B) COS7 cells were cotransfected with CFP-VSVG (red) and YFP-YFPAIA (green). (C) Scheme depicting the effects of BFA, nocodazole, and both, on ER export. BFA blocks ER export and causes COPI release from ERES membranes. Nocodazole causes the formation of Golgi mini-stacks adjacent to the ERESs. In the presence of nocodazole and BFA, cargo accumulates in the ERESs, which become dilated.





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