ROCK Inhibitor-Induced Promotion of Retinal Pigment Epithelial Cell Motility during Wound Healing

Figure 1

The effect of 10 μM Y27632 on RPE wound healing. (a) Histogram showing cell migration distances of RPE cells treated with and without Y24632, for each; . (b) The left figure represents phase-contrast image of untreated RPE cells at 0 hours after Y27632 administration. The middle and right figures represent automated visual-tracking of RPE cells treated with (right) and without (middle) Y24632 at 8 hours after Y27632 administration, for each. (c) Time course showing cell coverage percentage of RPE cells treated with and without Y24632 at 0, 4, 8, 12, 16, 20, and 24 hours after Y27632 administration. The solid line represents untreated RPE cells. The dashed line represents Y27632-treated RPE cells, for each; . (d) The left figure represents the open wound field between cells in the imported images, which were manually outlined. The middle and right figures represent phase-contrast images of RPE cells treated with (right) and without (middle) Y24632 at 24 hours after Y27632 administration. (e) F-actin (red), vinculin (green), and DAPI (blue) stained confocal images of wound-adjacent untreated RPE cells. (f) F-actin (red), vinculin (green), and DAPI (blue) stained confocal images of wound-adjacent Y24632-treated RPE cells. (g) F-actin (red), vinculin (green), and DAPI (blue) stained confocal images of untreated RPE cells far from wound sites. (h) F-actin (red), vinculin (green), and DAPI (blue) stained confocal images of Y24632-treated RPE cells far from wound sites. (i) Histogram showing cell coverage percentage of the ex vivo porcine RPE cells treated with and without Y24632 at 24 hours after Y27632 administration, for each; . (j) The autofluorescence images of porcine RPE-choroid-scleral fragment. RPE cells blocked scleral autofluorescence, and the scraped RPE area is represented as a green area. The figures represent the autofluorescence images of porcine RPE-choroid-scleral fragment before Y27632 administration (left) and treated with (right) and without (middle) Y24632 at 24 hours after Y27632 administration. (k) Hematoxylin-Eosin stained image of porcine RPE-choroid-scleral fragment. Black arrow represents the wound edge, and scraped RPE area is on the right side of black arrow.