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. Author manuscript; available in PMC: 2009 Nov 1.
Published in final edited form as: Arch Biochem Biophys. 2008 Aug 20;479(1):63–68. doi: 10.1016/j.abb.2008.08.009

Fig. 1. AP-1 forms higher-order macromolecular complexes upon Golgi recruitment.

Fig. 1

(A) Sucrose gradient fractionation of AP-1 recruited onto enriched Golgi membranes. 100 µg of rat liver Golgi membranes were incubated with 5 mg/ml bovine adrenal cytosol and 100 µM GTPγS for 15 min at 37 C in a 2 ml reaction. The membranes were collected by centrifugation and solubilized in assay buffer containing 2 % taurocholate. The lysate or 200 µl cytosol were loaded onto 10–25 % linear sucrose gradient and the fractions analyzed by SDS-PAGE and western blotting. The sedimentation coefficients (Svedberg units) of standard proteins run on an identical gradient are shown above. (B) Gel filtration analysis of Golgi-recruited AP-1. Golgi-recruited AP-1 was solubilized and analyzed by Superose 6 column chromatography as described in the Experimental Procedures.