Analysis of Bil1 localization and function in vivo. (A) Comparisons of Bud6-GFP localization in wild-type and bil1Δ cells (top), and Bil1-GFP localization in wild-type and bud6Δ cells (bottom). Bar, 10 µm. (B) Representative images of Bil1-GFP and Bud6-RFP localization in the same cells. Strong overlapping signals were observed at the bud neck (cyan arrowheads). Bar, 10 µm. (C) 10-fold serial dilutions of the indicated strains grown on YEPD plates at either 25 or 37°C. (D) Representative images of F-actin staining for the indicated strains. Bar, 5 µm. (E) Quantification of actin cable phenotypes (same categories as in Fig. 1, D and E). Note that data for some of the strains that appear in Fig. 1 D again appear here to facilitate comparison of phenotypes among relevant strains. The data for all of these strains were collected in the same sets of experiments. For each strain, >200 cells were scored (approximately equal numbers of cells pooled from two independent experiments). (F) Tracking of GFP-Sec4 particles in cells. Moving particles were followed over a 60-s observation window and placed into one of two categories: (1) normal anterograde movements (particles moving directionally toward the bud) or (2) random/retrograde movements. For each different yeast strain, two to three individual particles were tracked in 50 cells (n > 140 particles total; from approximately equal numbers of cells imaged in two independent experiments).