Figure 2. COPI budding from synthetic... | Rockefeller University Press

$Figure 2. COPI budding from synthetic liposomes and Golgi membranes analyzed by cryo-EM. (A and B) Cryo-EM of membranes incubated with Arf1-wt or Arf1-Y35A in the presence of the guanine nucleotide exchange factor ARNO, coatomer, and GTPγS. Images show liposomes (A) and Golgi membranes (B). Bars, 200 nm. (C) Purification of reconstituted of COPI vesicles. Recombinant Arf1-wt or Arf1-Y35A was incubated with Golgi membranes in the presence of GDP or GTP as in Fig. 2 B. Vesicles were purified by sucrose density centrifugation. Samples in the vesicle fraction were analyzed by SDS-PAGE and Western blotting using antibodies against the membrane marker transferrin receptor (TfR) and against coatomer subunit δ-COP and Arf1.$

Figure 2.

COPI budding from synthetic liposomes and Golgi membranes analyzed by cryo-EM. (A and B) Cryo-EM of membranes incubated with Arf1-wt or Arf1-Y35A in the presence of the guanine nucleotide exchange factor ARNO, coatomer, and GTPγS. Images show liposomes (A) and Golgi membranes (B). Bars, 200 nm. (C) Purification of reconstituted of COPI vesicles. Recombinant Arf1-wt or Arf1-Y35A was incubated with Golgi membranes in the presence of GDP or GTP as in Fig. 2 B. Vesicles were purified by sucrose density centrifugation. Samples in the vesicle fraction were analyzed by SDS-PAGE and Western blotting using antibodies against the membrane marker transferrin receptor (TfR) and against coatomer subunit δ-COP and Arf1.

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