FIGURE 3:

Bnr1-mediated actin cable assembly requires Gin4, Elm1, and Shs1. (A) Temperature-sensitive growth of gin4Δ bni1-1, elm1Δ bni1-1, and shs1Δ bni1-1 strains. Fivefold dilutions of cells were spotted on yeast extract/peptone/dextrose plates and grown for 1 d at 35°C (middle) or 37°C (bottom) or for 2 d at 24°C (top). (B) Myo2-GFP localization is depolarized in gin4Δ bni1-1, elm1Δ bni1-1, and shs1Δ bni1-1 after a 15-min shift to 35°C. Cells were imaged in three dimensions with five 0.5-μm stacks using the same exposure conditions for all experiments. Images are maximum-projection images; scale bar, 5 μm. (C) Quantification of Myo2-GFP localization from experiments in B. Cells of the indicated genotype were scored for Myo2-GFP localization to the bud tip (white with dots) or the bud neck (black) or unpolarized localization (gray). Experiments were performed three times (n > 200 cells for each repetition). For each double mutant, at least two independent clones from different spores were analyzed (data not shown).