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. 2001 Nov 12;155(4):557–570. doi: 10.1083/jcb.200107045

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Copyright © 2001, The Rockefeller University Press

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Figure 6. — The COPII coat actively cycles on and off the membrane in cells with disrupted ER–Golgi trafficking. (A) NRK cells stably expressing Sec13–YFP were transfected with Sar1[H79G]. They were fixed and then stained for either immunofluorescence or immunoEM with the HA epitope on the Sar1 construct. (B) Photobleaching of Sec13–YFP labeled structures in living cells. Cells were either left untreated, BFA-treated, or expressing Sar1[H79G] as indicated. The marked area was photobleached and fluorescence recovery was subsequently monitored. The arrows point to Sec13-labeled puncta before bleaching and following recovery. (C) Mean fluorescence intensity of the bleached region and nonbleached pools of Sec13–YFP were determined at the times indicated, and expressed as a percentage of the prebleach ratio between these values. (D) A model depicting the GTPase cycle of Sar1, indicating where the cycle is blocked in the mutants used in this study. Bars: (A, left) 5 μm; (A, right) 130 nm; (B) 5 μm.