DGK-α is required for pseudopod elongation evoked by inhibition of αvβ3 or expression of mutant p53. (A–D) A2780 cells (A and B) or vector and p53^273H-expressing H1299 cells (C and D) were transfected 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)) and plated onto CDM. Time-lapse videos were recorded in the absence (−) or presence of 36 nM cRGDvN^MeV or 2.5 µM cRGDfV in combination with 10 µM R59022 or its vehicle (DMSO) and 0.5% butan-1-ol (But-1-ol; or secondary alcohol control [0.5% butan-2-ol]). Representative stills are displayed. (A and C) The arrows indicate elongated pseudopods. Bars, 20 µm. In B and D, the distance between the center of the nucleus and the cell front (with respect to the direction of migration) was measured using ImageJ. Data are represented as box and whiskers plots (black lines show medians; whiskers: 5–95 percentile); n = 3 independent experiments. ***, P < 0.0001; Mann–Whitney test.