Expression of EVL is sufficient to promote mechanically directed motility and cell-matrix adhesion in MV^D7 cells. (a–d) GFP and GFP-EVL MV^D7 cells, plated on 35-kPa hydrogels, were mechanically stimulated. (a) Still images from representative time-lapse videos of GFP and GFP-EVL cells (Video 8). Scale bars are 10 µm. Boxed insets are images of expression constructs. (b) Corresponding cell traces at 0, 5, 10, 15, 20, 25, and 30 min, with starting positions in tan and final positions in blue. Crosshairs denote micropipette positions. (c) Rose plots show cumulative turning angles for GFP and GFP-EVL cells. Black sectors denote turns in the direction of the force gradient, and gray sectors denote turns away from the force gradient. Data are collected from five independent experiments (n ≥ 15 per condition). (d) Sensing indices of GFP and GFP-EVL cells over time. Two-way ANOVA shows a significant difference in sensing index between GFP and GFP-EVL cells (P < 0.0001). Data are collected from five independent experiments; all data points are shown (n ≥ 15 per condition; violin plot shows median and quartiles of sensing indices). (e) Top: Representative TIRF images of paxillin staining with GFP or GFP-EVL in MV^D7 cells. Boxed insets are inverted images of expression constructs. Bottom: Inverted single channel images of paxillin staining shown for clarity. Scale bars are 10 µm. (f) Quantification of FA area. Data are collected from three independent experiments; all data points are shown (n ≥ 121 per condition; P values were determined using regression analysis; ***, P ≤ 0.001; n.s., not significant; exact P values are found in Table S2; mean ± SEM). (g) Immunoblot of MV^D7 cells infected with control GFP vector or GFP-EVL. Top crop was probed with an antibody against EVL, and bottom crop was probed with an antibody against GAPDH.