Figure 4.
Vinculin-mediated phenotypes are ROCK-dependent. (A) Mean intensity of 2-NBDG for cells treated with 20 µM Y-27632 (Y27) or the vehicle control, and fold change of the 2-NBDG signal of cells treated with 20 µM Y-27632 relative to the vehicle control condition for each cell line (N = 3, n = 48–53 cells). (B) Representative images of 2-NBDG staining displayed using the Fire lookup table to indicate the relative intensity of the fluorescent signal. Scale bar, 50 µm. (C) Mean intensity of TMRM for cells treated with 20 µM Y-27632 or the vehicle control, and fold change of the TMRM signal of cells treated with 20 µM Y-27632 relative to the vehicle control condition for each cell line (N = 3, n = 95–118 cells). (D) Representative images of TMRM staining displayed using the Fire lookup table to indicate the relative intensity of the fluorescent signal. Scale bar, 50 µm. (E) Mean intensity of 2-NBDG for cells treated with 25 µM Blebb or the vehicle control, and fold change of the 2-NBDG signal of cells treated with 25 µM Blebb relative to the vehicle control condition for each cell line (N = 3, n = 80–88 cells). (F) Mean intensity of TMRM for cells treated with 25 µM Blebb or the vehicle control, and fold change of the TMRM signal of cells treated with 20 µM Blebb relative to the vehicle control condition for each cell line (N = 3, n = 76–84 cells). (G) Average area of FAs for cells treated with 20 µM Y-27632 or the vehicle control (N = 3, n = 46–50 cells). (H) Representative images of cells treated with 20 µM Y-27632 or the vehicle control and stained for paxillin (Pax), F-actin, and DAPI. Scale bar, 20 µm. Bar graphs denote the mean ± SEM. ns = not significant, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Vinculin-mediated phenotypes are ROCK-dependent. (A) Mean intensity of 2-NBDG for cells treated with 20 µM Y-27632 (Y27) or the vehicle control, and fold change of the 2-NBDG signal of cells treated with 20 µM Y-27632 relative to the vehicle control condition for each cell line (N = 3, n = 48–53 cells). (B) Representative images of 2-NBDG staining displayed using the Fire lookup table to indicate the relative intensity of the fluorescent signal. Scale bar, 50 µm. (C) Mean intensity of TMRM for cells treated with 20 µM Y-27632 or the vehicle control, and fold change of the TMRM signal of cells treated with 20 µM Y-27632 relative to the vehicle control condition for each cell line (N = 3, n = 95–118 cells). (D) Representative images of TMRM staining displayed using the Fire lookup table to indicate the relative intensity of the fluorescent signal. Scale bar, 50 µm. (E) Mean intensity of 2-NBDG for cells treated with 25 µM Blebb or the vehicle control, and fold change of the 2-NBDG signal of cells treated with 25 µM Blebb relative to the vehicle control condition for each cell line (N = 3, n = 80–88 cells). (F) Mean intensity of TMRM for cells treated with 25 µM Blebb or the vehicle control, and fold change of the TMRM signal of cells treated with 20 µM Blebb relative to the vehicle control condition for each cell line (N = 3, n = 76–84 cells). (G) Average area of FAs for cells treated with 20 µM Y-27632 or the vehicle control (N = 3, n = 46–50 cells). (H) Representative images of cells treated with 20 µM Y-27632 or the vehicle control and stained for paxillin (Pax), F-actin, and DAPI. Scale bar, 20 µm. Bar graphs denote the mean ± SEM. ns = not significant, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

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