Fig. 7
Measurements of diffusivity of exocyst subunits. a TIRFM images of untagged wild-type or indicated exocyst subunits fused to sfGFP (top). Heat map of above images (bottom). Scale bar = 20 μm. b Quantification of exocyst subunits localization at the TIRF field. Membrane localization index = density of spots * intensity of cells. Whiskers extend from min to max. Values for SEC3 and SEC6 were corrected for heterozygosity. n = 31, 13, 13, 13, 32 fields containing 2–4 cells each. c Particle tracking over time for the subunits indicated. For two-color tracking, each channel was tracked using Imaris software tracking algorithm and overlaid. Graphs show mean squared displacements over time. Scale bar = 0.5 μm (SEC3-GFP) and 0.8 μm (rest). d Representation of data in b as geometric means of MSDs < r2 >. e Diffusion coefficients of the indicated subunits were measured from < r2 > . Mean (µm2 s–1) ± SEM = 0.31 ± 0.05 (SEC3-GFP), 0.04 ± 0.002 (SEC5-Halo), 0.13 ± 0.01 (SEC6-GFP), 0.04 ± 0.004 (SEC8-GFP), and 0.05 ± 0.004 (EXO70-GFP). The effect size in terms of Cohen’s d value between SEC3-GFP and SEC6-GFP is 0.348 and between SEC6-GFP and SEC8-GFP is 0.176. White circles show the medians; box limits indicate the 25th and 75th percentiles; whiskers extend 1.5 times the interquartile range from the 25th and 75th percentiles; polygons represent density estimates of data and extend to extreme values. Experiments were repeated at least three times with similar results. P-values were computed using one-way ANOVA test followed by Scheffe’s multiple comparison tests
