Figure 7.

Vacuoles bearing Nyv1p-CCIIM require additional Sec18p and Vam7p for lipid mixing. A portion of BJ3505 NYV1-CCIIM vacuoles were labeled with self-quenching levels of octadecyl rhodamine B (R18), reisolated, and incubated with an unlabeled portion of these vacuoles under standard fusion conditions to assay for lipid mixing-induced dequenching, as described (Jun and Wickner, 2007). Measurements were taken every 2 min for 90 min, yielding fluorescence values at the onset (F₀) and during the reaction (F_t). The final 10 measurements of a sample containing 0.33% (v/v) Triton X-100 were averaged and used as a value for the fluorescence after infinite dilution (F_TX100). The relative total fluorescence change ΔF_t/F_TX100=(F_t−F₀)/F_TX100 was calculated.