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. 2009 Nov 15;20(22):4640–4651. doi: 10.1091/mbc.E09-05-0429

Figure 1.

Figure 1.

Syp1p and Ede1p dynamics at endocytic sites. (A) Epifluorescence images of live wild-type cells expressing Syp1-GFP or Ede1-GFP (strains DDY3865 and -3866). (B) Epifluorescence images of a single cell coexpressing Syp1-GFP and Ede1-RFP (strain DDY3871). (C) Dynamic cell surface localization of Syp1-GFP and Ede1-GFP (viewed by TIRF microscopy) in reference to Abp1-RFP (viewed by epifluorescence microscopy). Time series shows individual patches from two-color movies. Frame interval, 2 s. The proteins were coexpressed in strains DDY3867 and -3868. (D) Dynamic cell surface localization of Syp1-GFP and Ede1-GFP (viewed by TIRF microscopy) in reference to Sla1-mCherry (viewed by epifluorescence microscopy). Time series shows individual patches from two-color movies. Frame interval, 2 s. The proteins were coexpressed in DDY3899 and -3900. (E) Average lifetimes of Syp1-GFP and Ede1-GFP patches ± SD, n = 60. Data taken from 4-min TIRF microscopy movies with 2-s frame intervals. (F) Distribution of Syp1-GFP and Ede1-GFP patch lifetimes from the same dataset as in C. (G) Kymographs of representative Syp1-GFP and Ede1-GFP patches from epifluorescence movies. Patches were oriented so the outside of the cell is on top. (H) Quantification of fluorescence intensity for individual Syp1-GFP and Ede1-GFP patches over time. Each curve represents data from one patch. Fluorescence intensity was corrected for photobleaching. Movies were taken with 2-s frame intervals. All scale bars, 2 μm.