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Fig. 6. Cells expressing ^GFPRab11b mutants are not able to migrate efficiently in an experimental wound assay due to a directionality defect. Confluent cell monolayers were scratch-wounded to induce migration into the wound. Positions of cells approximately 60 minutes after wounding the monolayers (`start', A,D,G,J) and 6 hours later (`finish', B,E,H,K) are shown for the control PtK1 cells (A,B), cells stably expressing ^GFPRab11b-GDP (D,E) and ^GFPRab11b-GTP (G,H). The tracks of individual cells at the wound edges, as determined from time-lapse movies, are indicated in the `start' micrographs (A,D,G). Each frame constitutes a wound segment of approximately 400 µm in length. Graphs in panels C, F and I show X and Y positions of individual cells at the wound edge for the mutants in the micrographs above (the starting points of all cells are placed at 0:0). Data points are 2 hours apart. (J) Average translocation of the edge (i.e. total advancement into the wound) in 6 hours. (K) Mean average velocities of the mutant Rab11b-expressing cells at the wound edges and in the middle of the monolayers. Corresponding supplemental movies are available for control (supplementary material Movie 7), ^GFPRab11b-GDP (supplementary material Movie 8) and ^GFPRab11b-GTP (supplementary material Movie 9) cells.

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