Figure 1. P-cadherin expression induces... | Rockefeller University Press

Figure 1.

$Figure 1. P-cadherin expression induces CCM. (a) Protein extracts (20 µg/well) from the indicated cells were immunoblotted to detect E-, P-, R-, N-, and M-cadherin and β-actin. (b) Quantification of the indicated cadherins at the plasma membrane, normalized to the total amount of the corresponding cadherin, calculated from three independent experiments. (c and d) Persistence over 10 h after removal of the insert (c) and mean velocity and persistence measured between 4 and 15 h after removal of the insert (d). n = 242 C2C12 LZRS and 249 C2C12 Pcad cells from 15 independent experiments and 230 C2C12 Ecad and 171 C2C12 Rcad cells from 4 independent experiments. a.u., arbitrary units. (e) Trajectories over 15 h of 17 representative cells. Rose plots of angle trajectories (i.e., directionality). The magnitude of each bar shows the fraction of cells with the indicated angle trajectory. n = 193. (f) Migrating cells (8 h after insert removal) stained for nucleus, centrosome, and Golgi distribution. Arrows indicate the migration direction. (g) Histogram representing the percentage of migrating cells in which the centrosome and Golgi are located in the quadrant facing the free space in front of the nucleus (see cartoon) as an indication of cell polarization. n = 80 cells from five independent experiments. (h) Velocity fields and corresponding phase-contrast images and velocity vector orientation measured using MatPIV in the entire cell layer 10 h after insert removal. n = 983 C2C12 LZRS, 1006 C2C12 Pcad, 677 C2C12 Ecad, and 633 C2C12 Rcad. (i) Mean velocity measured in the entire cell layer from 4 to 15 h. All panels: means ± SEM. *, P < 0.05; **, P < 0.005; ***, P < 0.0005. Bars: (f) 15 µm; (h) 100 µm.$

P-cadherin expression induces CCM. (a) Protein extracts (20 µg/well) from the indicated cells were immunoblotted to detect E-, P-, R-, N-, and M-cadherin and β-actin. (b) Quantification of the indicated cadherins at the plasma membrane, normalized to the total amount of the corresponding cadherin, calculated from three independent experiments. (c and d) Persistence over 10 h after removal of the insert (c) and mean velocity and persistence measured between 4 and 15 h after removal of the insert (d). n = 242 C2C12 LZRS and 249 C2C12 Pcad cells from 15 independent experiments and 230 C2C12 Ecad and 171 C2C12 Rcad cells from 4 independent experiments. a.u., arbitrary units. (e) Trajectories over 15 h of 17 representative cells. Rose plots of angle trajectories (i.e., directionality). The magnitude of each bar shows the fraction of cells with the indicated angle trajectory. n = 193. (f) Migrating cells (8 h after insert removal) stained for nucleus, centrosome, and Golgi distribution. Arrows indicate the migration direction. (g) Histogram representing the percentage of migrating cells in which the centrosome and Golgi are located in the quadrant facing the free space in front of the nucleus (see cartoon) as an indication of cell polarization. n = 80 cells from five independent experiments. (h) Velocity fields and corresponding phase-contrast images and velocity vector orientation measured using MatPIV in the entire cell layer 10 h after insert removal. n = 983 C2C12 LZRS, 1006 C2C12 Pcad, 677 C2C12 Ecad, and 633 C2C12 Rcad. (i) Mean velocity measured in the entire cell layer from 4 to 15 h. All panels: means ± SEM. *, P < 0.05; **, P < 0.005; ***, P < 0.0005. Bars: (f) 15 µm; (h) 100 µm.