Effects of tumor-expressed FVII expression on cell behavior. (A) Strategy for creating FVII-expressing cells. An FRT site was inserted into the genome of MDA-MB-231 breast cancer cells. Subsequently, the gene of interest (empty vector or FVII cDNA) was inserted into the FRT site by cotransfecting a recombinase-expressing construct. Subsequently, colocalization of FVII and TF (B), coagulant activity in the presence or absence of a FVII-blocking antibody (n = 3) (C), and FVII target gene expression (n = 5) were determined in MDA^pcDNA and MDA^FVII cells (D-E). (F) Invasion of MDA^pcDNA and MDA^FVII cells through Matrigel-coated transwell inserts in the presence of the indicated blocking antibodies (n = 4). (G) FVII-dependent gene expression in MDA-MB-453 cells after FVII knockdown by shRNA (n = 3). (H) Morphology of MDA^FRT, MDA^pcDNA, and MDA^FVII cells. Scale bars, 10 μm (B) 40 μm (H). Note the epithelial patches of FRT and pcDNA cells. All graphs show the mean and standard deviation (SD). Statistically significant differences were tested using t tests. IgG, immunoglobulin G.