FIGURE 5:

Eps15 can function as an endocytic-site initiator in the absence of Ede1. (A) Representative still images of cells expressing the Eps15-GFP using the indicated promoter. (B) GFP constructs from A were expressed with Sla1-mCherry and imaged for 4 min at 2 s/frame with ∼300-ms delay between channels. The first frame of a movie of a representative cell, followed by a circular kymograph of the bud and mother cortex. Scale bars, 2 μm and 40 s. (C) The patch traces identified in kymographs from B in two 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. The mean percentage and SD of each category per cell is indicated. (D) Quantification of kymograph analysis from two independent experiments of the number of patches that start during a 90-s movie in cells of the indicated genotype. The p values were calculated using Student’s t test. (E) Quantification of the Sla1-mCherry lifetime from two independent experiments in cells of the indicated genotype. (F) Quantification of kymograph analysis of the proportion of Sla1-mCherry patches that internalize from two independent experiments per cell (25 cells/strain).