DGK-α is necessary for RCP recruitment to pseudopod tips during migration on CDM. (A and B) A2780 cells (A) or vector and p53^273H-expressing H1299 cells (B) were transfected with GFP-RCP in combination with nontargeting siRNA (siRNA control [si-con]), an siRNA targeting DGK-α (si-DGKα), or siRNA targeting DGK-α in combination with siRNA-resistant DGK-α (si-DGKα/DGKα(resc)). Cells were plated onto CDM, mounted onto the stage of a confocal microscope (FV1000), and incubated in the absence (−) or presence of 36 nM cRGDfN^MeV or 2.5 µM cRGDfV. Images were captured at one frame/second over a period of 60 s, and videos were generated from these (Videos 4, 5, 6, and 7). Single-section confocal image stills corresponding to individual frames from these videos are presented. The arrows indicate the direction of migration, and the portion of the cell within the white square is presented in the bottom images. Bars, 10 µm.