Dia1 is dispensable for HGF-mediated planar motility in 2D and within acini. (A) Transmitted light images at 0, 5, and 9 h of shControl and shDia1 cell island scattering after 20 ng/ml HGF addition at 0 h. Time is indicated in h:min. See also Video 2. (B) Rose plots of 10 cell trajectories from the cell islands scattering in A and Video 2 over 9 h. The initial location of each trajectory was positioned at 0, 0. (C) Box plots of mean instantaneous speed and persistence for shControl and shDia1 cells. (D) Acini of shControl- and shDia1-expressing H2B-mCherry. Acini were stimulated with 20 ng/ml HGF and imaged by time-lapse fluorescence microscopy for 12 h. Shown are stills with individual tracks overlaid. See also Video 3. (E) Rose plots from cell trajectories obtained from D and Video 3. (F) Box plots of mean instantaneous speed and persistence to characterize cell motility, with the number of cell trajectories obtained from at least five acini per condition indicated below each plot. Box plots show the 25th and 75th percentiles and the median, circles indicate means, and whiskers mark 1.5 SDs. *, P < 0.01; **, P < 0.05 by a Student’s two-tailed t test assuming unequal variance.