Fig. 5.

Content mixing of RPLs bearing Nyv1p-CCIIM. In a complete fusion reaction with all the SNAREs and chaperones, content mixing was monitored by the FRET signal between streptavidin-Cy5 (from R-SNARE RPLs) and biotin-R-phycoerythrin (from Q-SNARE RPLs) with a large molar excess of biotin-dextran in the reaction (light gray columns). Content mixing plus lysis was monitored in a parallel reaction without biotin-dextran (dark columns). Where indicated, Vam7p was replaced with buffer or detergent was added. The maximal FRET signal in the absence of biotin-dextran was determined at t = 60 min. The FRET signals at 45 min as a percentage of the maximal signal from three independent experiments were averaged and used to compare the content mixing or lysis under different fusion conditions. Error bars represent SDs.