FIGURE 2:
A construct containing the coiled-coil domain, third EH domain, and PP region is sufficient to recruit coat proteins. (A) Lifetimes of individual Ede1 patches from cells of the indicated genotype calculated from three independent experiments (EDE1-GFP, 328 patches; ede1EH3PPCConly-GFP, 480 patches). Mean ± SD. The p values were calculated using the Student’s t test. (B) Ede1-GFP variants were imaged with Sla1-mCherry for 4 min at 2 s/frame using simultaneous dual-color imaging. The first frame of a movie of a representative cell, along with circular kymographs of the mother and bud cortex. Scale bars, 2 μm and 40 s. (C) The patch traces identified in kymographs from three independent experiments were segregated into one of four categories depending on the relative arrival of GFP and mCherry: GFP or mCherry alone, GFP followed by mCherry, or mCherry followed by GFP. Mean percentage and SD of each category per cell. (D) Ede1-GFP variants were imaged with Sla2-mCherry for 4 min at 2 s/frame using simultaneous dual-color imaging. The first frame of a movie of a representative cell is shown along with circular kymographs of the mother and bud cortex. Scale bars, 2 μm and 40 s.