Figure 5.

The early module regulates endocytic site formation and distribution. (A) Maximum intensity projections of Z-stacks of wild-type, syp1Δ, ede1Δ, and syp1Δ ede1Δ cells expressing Sla1-GFP (strains DDY3700, -3798, -3875, and -3876). Z-stacks were acquired through the entire cell at 0.15-μm intervals. Scale bar, 2 μm. (B) Patch number per cell surface area (μm²) in wild-type, syp1Δ, ede1Δ, and syp1Δ ede1Δ cells expressing the indicated GFP-tagged markers (strains DDY2736, -3696, -3697, -3700, -3701, -3798, and -3872–3885). n = 20 cells for each strain; error bars, SD. Patches were counted in approximately spherical unbudded or large-budded cells; cell surface area was estimated as an average of sphere surface areas calculated from four diameters measured from the maximum intensity projections. (C) Maximum intensity projections of Z-stacks of small-budded wild-type, syp1Δ, ede1Δ, and syp1Δ ede1Δ cells expressing the indicated GFP-tagged markers (strains DDY2736, -3696, -3697, -3700, -3701, -3798, and -3872–3889). Z-stacks were acquired through the entire cell with 0.15-μm intervals. Arrowheads indicate the bud neck region affected by SYP1 deletion. Scale bar, 2 μm. (D) Percentage of wild-type, syp1Δ, ede1Δ, and syp1Δ ede1Δ cells expressing the indicated GFP-tagged markers with patches polarized to the small-budded neck (using the same strains listed in C). Mean averages from three separate experiments are shown (n = 60 cells for each strain in each experiment; error bars, SD). Cells were scored as described in the text.