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. 2001 Dec 10;155(6):969–978. doi: 10.1083/jcb.200104093

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Copyright © 2001, The Rockefeller University Press

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Figure 3. — Characterization of in vitro TGN membrane fusion. (A) Processing of Ste13αHA requires an ATP regeneration system. Control reactions (±Kex2p) included 1.5 mM exogenous Mg-ATP, 40 mM phosphocreatine, and 0.125 mg ml⁻¹ creatine kinase. Omission of Mg-ATP (−ATP), or phosphocreatine/creatine kinase, (−Regen) is indicated. (B) Temperature dependence of Ste13αHA processing in vitro. Reactions were performed at each temperature using lysates from either KEX2 or kex2Δ strains. (C) Dilution sensitivity of Ste13αHA processing. Reactions were diluted to the indicated extents before (Pre-reaction) or after (Post-reaction) incubation under reaction conditions. (D) Detergent sensitivity. MSS membranes from either KEX2 or kex2Δ strains were combined with MSS membranes from a Ste13αHA-expressing strain, and reactions were performed in the presence or absence of 1% Triton X-100.