Fig. 8. Cell-cell contact loss from the effects of OXO on peripheral actin filaments is correlated with wound healing abilities that are dependent on integrin 5, PKC, SFK and Akt/PKB activities. (A) Confluent RIE1 cells seeded onto 60 mm culture dishes were wounded, washed twice with 10% FBS-DMEM-H, and treated with DMSO or 10 µM OXO, as described in Materials and Methods. Phase contrast images were taken 17 or 24 hours after wounding and treatment. (B) Confluent cells in 60 mm culture dishes were wounded, washed, and then pretreated with LY294002 (LY, 20 µM), PP2 (10 µM), PP3 (10 µM) or ML9 (20 µM) 10 minutes before 10 µM OXO treatment. Another set of cells was treated with DMSO alone in parallel. After incubation for 20 hours, phase contrast images were taken. Note that ML9 treatment caused a certain level of cytotoxicity. (C) Cells in 60 mm culture dishes were infected with adenovirus for GFP (Ad-cont), K376A DN PKC (Ad-DN PKC), or WT PKC (Ad-WT PKC) for 8 hours. After the viruses had been washed out, the cells were treated with DMSO or 10 µM OXO for 16 hours, and then images were taken of the wound area. Dotted lines indicate the starting lines for wound healing. Data shown are representative of three independent experiments.