FHOD3 is required for invasive migration in vitro and in vivo. (A) A2780 cells stably expressing pLVTHM vector control, shFHOD3 #1, or shFHOD3 #2 were seeded into inverted invasion assays and allowed to invade into Collagen-I/FN matrix for 72 h. Cells were visualized with calcein-AM and serial confocal sections were captured at 15-µm intervals. (B) Invasion of H1299-p53-273H cells stably expressing pLVTHM vector control, shFHOD3 #1, or shFHOD3 #2 was determined as in A. (C) A2780 cells stably expressing pLVTHM vector control, shFHOD3 #1, or shFHOD3 #2 were injected into the pericardial cavity of zebrafish embryos and treated with cRGDfV (2.5 µM). White arrows indicate invading cells. The broken lines indicate the pericardial cavity. (D) Relative invasion quantification of C. n = 24–27 fish per condition. All data represent at least three independent experiments. Statistical significance was evaluated using ANOVA/Tukey’s multiple comparison test. +, mean; *, P < 0.05; **, P < 0.01; ****, P < 0.0001.