Fig. 6. Phenotype of for3 null cells. (A) Viability of for3 null cells. Indicated cells were grown at 25°C for 4 days on a YEA plate. (B) Aberrant morphology of for3 null cells and rho3 for3 double null cells. Cells grown in YEA medium at 25°C. DIC images are shown. (C) Defects in actin cytoskeleton and MTs in for3 null cells. for3 null cells grown in YEA medium at 25°C were stained with Bodipy-phallacidin (green), TAT-1 (red), and DAPI (blue). Deconvoluted 3D images are shown. Small arrowhead indicates disorganized cytoplasmic MTs in dumpy cells. Large arrowhead indicates curved MTs located on a convex side of a bent cell. Arrows indicate F-actin cables. (D) Interphase F-actin cables in for3 null cells. for3 null cells grown in YEA medium at 25°C were stained with Bodipy-phallacidin and DAPI. The pictures represent deconvoluted Z-axis sections (top, middle, and bottom sections from left to right) of three interphase cells. Arrows indicate F-actin cables. (E) Formation of thick F-actin cables by overexpression of Fim1A2. Wild-type cells or for3 null cells containing pREP1 fim1 A2 were grown in EMM without thiamine for 18 hours at 25°C. F-actin images are shown. (F) Observation of cells expressing For3-truncated proteins. (a) Deconvoluted 3D images of for3 FH12C cells, for3 FH2C cells, for3 C cells, and for3 null cells containing pREP1 for3NC, pREP1 for3MC, pREP1 for3FH12, or pREP1 for3C are shown. The cells were stained as in C. (b) F-actin images of a for3 null cell containing pREP1 for3MC indicated by the large arrow in a. The pictures represent deconvoluted Z-axis sections (top, middle, and bottom sections from left to right). (G) A for3 rho3 null cell grown in YEA medium at 25°C was stained with Bodipy-phallacidin (left), and TAT-1 (right). Deconvoluted 3D images are shown. Bars, 5 µm.

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