Inhibition of αvβ3 promotes the formation of dynamic actin-rich protrusions at the cell front. (A) Confluent monolayers were wounded with a plastic pipette tip and the were cells allowed to migrate for 2 h into the wound in the absence (BASAL) or presence of 1 μM cilengitide (CIL) or 0.5 μg/ml osteopontin (OSP). Cells were fixed and permeabilized and F-actin was visualized using fluorescently conjugated phalloidin. 3D reconstructions were generated from serial Z sections captured using a confocal microscope. Yellow arrow denotes the direction of cell migration. Bar, 10 μm. Time-lapse movies indicating the dynamics of GFP-actin under conditions corresponding to the experiments presented in A are included as Videos 4–6, respectively . (B) A2780 cells were transfected with GFP-RCP^wt, GFP-RCP^621E, GFP-FIP2^480E, or GFP-Rip11^630E. Confluent monolayers were wounded with a plastic pipette tip and the cells were allowed to migrate for 2 h into the wound in the absence or presence of 1 μM cilengitide. Cells were fixed and permeabilized and GFP and F-actin was visualized by fluorescence microscopy. 3D reconstructions were generated from serial Z sections captured using a confocal microscope. Yellow arrow denotes the direction of cell migration. Bar, 10 μm.