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. 2002 Oct 14;159(1):69–78. doi: 10.1083/jcb.200206015

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Copyright © 2002, The Rockefeller University Press

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Figure 6. — Ultrastructural characterization of vesicles for GAP and ARF1. (A) Vesicles reconstituted in the presence of GTP contain detectable levels of GAP but not ARF1. The two-stage incubation was performed with the second stage using either GAP or palmitoyl-CoA, as indicated. A fraction (10% by volume) of the supernatant derived from the second-stage incubation was then applied onto grids for immunogold EM using antibodies as indicated. Magnification, 120,000. Bar, 50 nm. (B) The level of GAP on COPI vesicles is reduced when reconstitution is performed in the presence of GTPγS. The two-stage incubation was performed with the first stage using either GTP or GTPγ, and the second stage using GAP. A fraction (10% by volume) of the supernatant derived from the second stage was then applied onto grids for immunogold EM with antibody against GAP, followed by quantitation for vesicles with gold particles.

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