Defining molecular components of the early and late steps of the endocytic pathway. (A) Maximum-intensity projections of epifluorescence z-stacks of clusters of cells endogenously expressing Sla1-mCherry (magenta) and Syp1-GFP (green, first row), Sla2-GFP (green, second row), Ent2-GFP (green, third row), and Pan1-GFP (green, last row). Individual channels are shown in grayscale at left. (B) Quantification of the ratio of the number of Sla1-mCherry sites to the number of Syp1-GFP (top row), Sla2-GFP (second row), Ent2-GFP (third row), and Pan1-GFP (last row) sites from maximum-intensity projections of z-stacks of small-budded (polarized) and large-budded (depolarized) mother cells (n = 30 cells per category). Two-tailed P values from Mann–Whitney U tests as in Fig. 7 C (U_Syp1 = 0, U_Sla2 = 0, U_Ent2 = 0, U_Pan1 = 448) are displayed. The median and interquartile ranges are denoted with error bars. (C) Maximum-intensity projections of epifluorescence z-stacks of clusters of 7Δ cells endogenously expressing Sla1-mCherry (magenta) and Sla2-GFP (green). Individual channels are shown in grayscale at left. Yellow arrowheads highlight low concentration of Sla1-mCherry sites in a small bud, where late endocytic sites would normally be plentiful. (D) Quantification of the ratio of the number of Sla1-mCherry sites to the number of Sla2-GFP sites in small-budded (Sm) and large-budded (Lg) wild-type and 7Δ cells (n = 30 cells per category). Numbers are P values from Kruskal–Wallis tests followed by Dunn’s multiple comparisons test. The median and interquartile ranges are denoted with error bars.