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Fig. 2. Actin organisation in formin mutants. Analysis of F-actin structures in fixed cells of asynchronous populations stained by rhodamine-phalloidin. (A) Representative images for actin organisation in the indicated strains. Bar, 2 µm. (B) Distribution of budded cells from asynchronous populations (excluding cells at cytokinesis), according to the presence of polarised actin cables and patches, as depicted in the diagrams: thick or numerous cables directed towards the neck and polarised patches (1), thin or few cables directed towards the neck and polarised patches (2), disorganised cables but polarised patches (3), no visible cable but polarised patches (4), no visible cable and depolarised patches (5). Over 400 cells were scored. Error bars indicate s.e.m. (C,D) Bni1^CTp significantly supports cell polarity. Diploid budding pattern scored as an indirect indicator of actin organisation and cell polarity in wild-type cells or the indicated mutants. (C) Representative images of fixed diploid cells stained with calcofluor to label chitin-containing scars illustrating bipolar (left) or random (right) budding pattern. Bar, 2 µm. (D) Distribution of diploid cells according to their budding pattern (n=500 cells). bni1/BNI1 and bni1/bni1^CT exhibited comparable bipolar budding relative to a bni1/bni1. Error bars indicate s.e.m. (E,F) Analysis of actin organisation for synergism effects. (E) Synergistic loss of actin organisation and cell polarity in bud6 bni1 cells. (F) Synergism is not apparent between bud6 and bni1^CT bnr1 mutations. Cells were scored as indicated in B.

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