FIG. 5.

ARF proteins are partially lost from the cytoplasmic extract through treatment with GTPγS. Extract was divided evenly and either treated with indicated concentrations of GTPγS or mock treated as described in Materials and Methods. Following dialysis and centrifugation, equal aliquots of the soluble and insoluble fractions of all four extracts were analyzed by SDS-PAGE (15% gel) and ECL immunoblotting with anti-ARF monoclonal antibody. Quantitation of resulting signal was carried out by densitometry scanning of short-exposure films of the immunoblot and is presented as a percentage of ARF found in the supernatant fraction of the mock-treated extract.