Figure 4.

Localization interdependence of Syp1p and Ede1p. (A) Epifluorescence images of wild-type and ede1Δ cells expressing Syp1-GFP (strains DDY3865 and -3869). (B) Dynamic surface localization of Syp1-GFP in wild-type and ede1Δ cells viewed by TIRF microscopy. Left, single frames from movies; Right, kymograph representations of the same movies. Movies were taken with 1-s frame intervals for 2 min. (C) Epifluorescence images of small- (top), medium- (middle), and large-budded (bottom) wild-type and syp1Δ cells expressing Ede1-GFP (strains DDY3866 and -3870). Arrowheads, the bud neck region affected by SYP1 deletion. (D) Cell cycle distribution of Syp1-GFP and Ede1-RFP in wild-type cells (strain DDY3871). Individual cells were imaged every 10 min for 2 h, and one image is shown for every 20 min (the full sequence can be seen in Supplemental Movie S2); green and red images were acquired immediately after one another. White arrowheads, the first signs of neck localization during bud emergence (a second cell cycle has begun at 120 min); red arrowheads, localization to the bud neck at cytokinesis. Scale bars, 2 μm throughout.