Fig. 4. Time-lapse analysis reveals increased directionality of migration and impaired intercellular adhesion in AF6i3-knockdown cells. (A) Time-lapse images showing the wound closure of AF6i3-knockdown and control cells at the time of wounding (0 h) and 10 hours later (see also supplementary material Movies 1 and 2). Red lines mark the boundary of the wound at time point 0 seconds. Bar, 0.1 mm. (B) Representative trajectories of cells during wound healing assay shown in A, followed from time 0 for 10 hours. Bar, 0.1 mm. (C) Graphic representation of the directionality of migration, defined by the D/T ratio, which represents the direct distance from the cell track start to the end point (D) divided by the total path length (T). Data are derived from three independent experiments. (D) Wound margin of migrating AF6i3-knockdown and control cells 10 hours after wounding. green, anti-E-cadherin antibody; red, phalloidin-TRITC. Bar, 10 µm. (E) Western blot analysis of whole-cell lysates during wound healing assay.