FIGURE 4: The effect of Vps1 mutations on individual endocytic events. Cells expressing Sac6-mRFP and deficient for vps1 were transformed with an empty plasmid and wild type or mutant vps1. (A) Time-lapse movies were recorded and used to measure the lifetime of the reporter at endocytic sites. Means are reported, error bars are standard deviation. A 1-way-Anova was used with Dunnett’s multiple comparison test to assess significance of differences from the wild type. Those marked with asterisks are considered significant p≤0.0001. (B) Movies were also used to generate kymographs for each of the strains to analyze the behavior of patches during invagination. Indicated with arrows: R – retraction event, D- delayed scission/disassembly event, 2P – second patch event. (C) Patch tracks were generated to follow the movement of patches. Three are shown for each strain with the center of each spot marking the recorded position at each time point. Green spot indicates the start point and red the last point before disassembly.

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