Deletion of the Hof1 F-BAR domain in vivo gives rise to actin cable defects. (A) SIM of F-actin organization in fixed wild-type, hof1Δ, and hof1ΔFBAR cells. (B) Automated analysis of actin cable organization from SIM images as in A with SOAX, showing cable segments (purple) and cable intersections (green). (C) Polarity map of actin cable segments in representative cells from the SIM images, with color-coding according to azimuthal orientation of cable segments relative to mother-bud axis. Azimuthal angle is defined such that the angle is zero when a cable segment is parallel with the x axis with a range of ±90 degrees. Cells analyzed marked with a yellow arrowhead in B. (D) Average number of cable segments per cell analyzed by SOAX, quantified for 20 cells per strain. (E) The same cells were quantified for number of cable intersections per cell by SOAX. (F) CVs of F-actin staining in mother cells after treatment with 100 µM CK666. Data were averaged for 20 cells in each strain. (G) Representative traces (18 for each strain) of GFP-Sec4 paths, tracking vesicle movement from its starting position in the mother cell until it reached the bud neck. Convergence point of traces indicates the bud neck. (H) Quantification of path lengths of GPF-Sec4 vesicle movements, imaged as above (n = 100 vesicles per strain). (I) Tortuosity (length/distance) of the paths of the same vesicles. (J) Quantification of actin cable extension velocities from live-imaging experiments using Abp140-3xGFP to decorate cables (n = 25 cables per strain). Error bars in all panels represent SEM. Statistical significance in all panels was calculated by one-way ANOVA (n.s., not significant; *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001).