= Solution
Cold preincubation can accumulate docked, fusion-ready intermediates. On warming, that population proceeds promptly to lipid mixing, whereas newly combined vesicles must first encounter, dock and assemble productive <SNARE complexes>. This explains the faster initial rise and larger signal at the displayed endpoint without requiring a different total amount of fluorescent lipid.
The original reconstitution experiments likewise distinguish docking from subsequent <membrane fusion>.<SNAREpins: minimal machinery for membrane fusion by T. Weber and others>[Docked unfused intermediates accumulated at low temperature and fused on warming]{ref}
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