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Figure 3


Fig. 3. Munc18-1 prevents ectopic t-SNARE interactions. (A) Wild-type or mutants of Syx1-288 (red), SNAP-25 (green) and munc18-1 were coexpressed in live HEK293 cells and imaged by CLSM. The merged image shows areas of coincidence in yellow hues. The two-dimensional histogram represents the intensity of each channel in each voxel, with a colour scale representing frequency. The residual map corresponds to weighted residuals from the line fit to the histogram, thus indicating fluorescent channel covariance. The hue is from –1 to 1, with cyan corresponding to a zero residual. (B) Similar results were obtained using N2a cells. Bars, 5 µm. (C) The covariance data from HEK293 cells coexpressing wild-type or mutants of Syx and SNAP251-206, in the absence (filled bars) or presence of munc18-1 (open bars) were quantified and expressed as Pearson's coefficient values. These data showed that the covariance between the t-SNAREs is unaffected by the presence of munc18-1, but that the location of the colocalisation is altered (see panel A; n>=4). Data are expressed as the mean±s.e.m.





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