Fig. 4. Bud6^1-565p restored microtubule capture but not actin organisation to a bud6 strain. (A) Representative images for localisation of GFP-Bud6p or GFP-Bud6^1-565p expressed under the HIS3 promoter in bud6 cells. GFP-Bud6^1-565p was efficiently localised to the bud and bud neck. Images are 2D projections of five-plane z-stacks. Bar, 2 µm. (B,C) Actin organisation defects of bud6 were not rescued by expression of Bud6^1-565p. (B) Representative images of fixed cells stained with rhodamine-phalloidin showing actin organisation in bud6 cells expressing Bud6p or Bud6^1-565p. Bar, 2 µm. (C) Quantitation of actin organisation for the indicated strains as in Fig. 2; n>400 cells. Error bars indicate s.e.m. (D) MT–cell cortex interactions scored in time lapse series of the indicated strains during the interval from mitotic exit to bud emergence as in Fig. 4C. At least 140 cortical interactions were scored per strain. Expression of full-length Bud6p or Bud6^1-565p restored MT shrinkage in a bud6 mutant (P<0.0001) to the level observed in wild-type cells. (E) Correct preanaphase spindle orientation along the mother bud axis was scored (Theesfeld et al., 1999; Huisman et al., 2004) in the indicated strains showing that bud6^1-565 suppressed the genetic interaction between kar9 and bud6. (F) Spindle polarity in anaphase cells (only one SPB in contact with the bud) (Huisman et al., 2004) was scored in the indicated strains, showing that bud6^1-565 suppressed the genetic interaction between dyn1 and bud6.