Figure 4. Rho A pulldown assay, ROCK activity assay, and myosin light chain phosphorylation in TrypLE-prepared and collagenase-A-prepared BCECs. In the TrypLE-prepared bovine corneal endothelial cells (BCECs), the Y-27632 treatment reduced Rho A activation on days 1, 3, and 5. Rho A activation decreased at confluence status on day 7 (A). In the collagenase-A-prepared BCECs, Y-27632 treatment reduced Rho A activation on days 1, 3, 5, and 7 (confluence status; B). In the TrypLE-prepared BCECs, the Y-27632 treatment reduced ROCK activity on days 1, 3, and 5 but exhibited no inhibitory effects at confluence on day 7 (C). In the collagenase-A-prepared BCECs, the Y-27632 treatment inhibited ROCK activity on days 1, 3, 5, and 7 (confluence status; D). The Y-27632 treatment reduced the amount of phosphorylated myosin light chain in the TrypLE-prepared BCECs on days 3, 5, and 7 (E), and reduced the amount of phosphorylated myosin light chain in the collagenase-A-prepared BCECs on days 1, 3, 5, and 7 (F). * indicates p<0.05.