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. 2015 Jul 1;26(13):2519–2534. doi: 10.1091/mbc.E14-11-1527

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© 2015 Jose et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 5: — Endocytic and exocytic vesicles become interspersed in exocyst mutants. (A) Steady-state image of a fixed control cell. The maximum intensity projection of a deconvolved 3D image stack. Scale bar, 2 μm. Bottom, fluorescence distribution (y-axis) of the endocytic (red) and exocytic (cyan) compartments along the bud–mother axis (x-axis, from left to right) in small-budded cells. Error bars denote SD. An asterisk indicates the position of the average bud neck. (B) As in A, but showing a sec6-4 mutant after shift to the restrictive temperature. (C) Immunoblot analysis of the levels of GFP-Sec4 at 25 and 34°C in untagged cells, sec6-4 cells, and an isogenic wild-type control. The protein Rna15 was detected as a loading control. (D) Simultaneous, dual-color near-TIRFM to examine in vivo dynamics of endocytic (red) and exocytic (cyan) vesicles in the sec6-4 exocyst mutant at 25 and 34°C. Cells were grown at 25ºC and then imaged at the indicated temperature. The dashed lines represent the region selected for generating the kymograph. The kymograph on the left shows the merge of the endocytic and exocytic channels; the right kymograph shows the endocytic channel alone.