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. 2015 Feb 25;290(16):10535–10543. doi: 10.1074/jbc.M114.631341

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© 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 3. — Munc18-1 and Syt1 are mutually antagonistic in SNARE-dependent lipid mixing. A, normalized distributions of FRET efficiencies for immobilized t-vesicles and docked VAMP2-vesicles (gray), VAMP2-vesicles in the presence of 10 μm Vps (red), VAMP2-vesicles in the presence of 1 μm Munc18-1 (blue), and VAMP2-vesicles in the presence of 10 μm Vps and 1 μm Munc18-1 (dark cyan). B, normalized distributions of FRET efficiencies for immobilized t-vesicles and docked VAMP2/Syt1-vesicles (gray), VAMP2/Syt1-vesicles in the presence of 10 μm Vps (red), VAMP2/Syt1-vesicles in the presence of 1 μm Munc18-1 (blue), and VAMP2/Syt1-vesicles in the presence of 10 μm Vps and 1 μm Munc18-1 (dark cyan). C, lipid-mixing efficiencies quantified based on the percentages of normalized high FRET population (E ∼0.5–1) (65). Bars are normalized with respect to the number of the docked vesicles to take into account the differences in docking probabilities (D). E, lipid-mixing efficiencies quantified based on the percentages of the normalized high FRET population (E ∼0.5–1) (65) for the t-vesicles without PIP₂. Bars are normalized with respect to the number of the docked vesicles (F). Error bars, S.D. of three independent experiments; movies were recorded for analysis from more than five randomly selected screens in each experiment. *, p < 0.05, assessed using the two-sample t test, unless otherwise specified.