Cilengitide drives RCP to the tips of extending pseudopods. (A and B) Cells were transfected with GFP-RCP^wt or GFP-RCP^621E and plated onto cell-derived matrix in the presence and absence of 1 μM cilengitide ∼4 h before time-lapse microscopy. Images were captured every 5 min over a 6-h period and movies were generated from these (Videos 7–9) and stills from these movies are presented. Bar, 100 μm. (B) Speed and persistence of migration were determined as for Fig. 3 B, values are mean ± SEM; n = 3 independent experiments. To obtain a measure of pseudopod length, the distance between the center of the nucleus and the cell front (with respect to the direction of migration) was measured using ImageJ. Data represents mean ± SEM; n = 3 independent experiments. (C) Cells were transfected with GFP-RCP, GFP-RCP^621E, or GFP-RCP^379-649 and plated onto cell-derived matrix in the presence and absence of 1 μM cilengitide and imaged by confocal microscopy. Images were captured at 1 frame/second over a period of 100 s and movies were generated from these. Single section confocal image stills corresponding to individual frames from these movies are presented. Bar, 20 μm. The yellow arrows indicate the direction of migration and the portion of the cell within the white square is presented as Video 10. Videos are available.