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Fig. 8. Sec31B-F moves with COPII complexes at different temperature or after disruption with nocodazole. The MDCK-787 cells expressing eYFP-Sec31B-F were transfected with VSV-G, and the dynamics of these proteins was measured under conditions of low-temperature blockage of ER export, or nocodazole disruption of microtubule-based intracellular transport. (A). Cells blocked at 15°C were warmed to room temperature (RT) for 4 minutes to allow entry of VSV-G into the ERESs. Cells were then fixed and examined for the above proteins. Under these conditions, VSV-G clusters into hundreds of minute ERESs that are coincident with COPII vesicular tubular clusters as detected by their Sec13 staining. The distribution of eYFP-Sec31B-F was perfectly coincident with these COPII-enriched ERESs. (B) Similar preparation of MDCK-787 cells blocked with nocodazole and incubated for 15 minutes at RT after a 2 hour incubation at 40°C. Under these conditions, eYFP-Sec31B-F forms coarse puncated clustes closely associated with the dispersed Golgi network to which VSV-G had moved during the 15 minute incubation at the permissive temperature (RT). Bars, 10 µm.
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