Fig. 6. Dynamic reorganization of peripheral actin filament and cell-cell contact loss of RIE1-5 cells depend on PKC-dependent cofilin dephosphorylation after OXO treatment. (A-C) Cells in 60 mm culture dishes were infected with adenovirus for GFP (Cont), K376A DN PKC (DN), or WT PKC (WT), for 8 hours (A). Cells were infected or transiently transfected with adenovirus for K368R DN or WT PKC (B) or K437R or WT PKC-HA (C), respectively. (D-I) Cells on 10% FBS-DMEM-H-coated glass coverslips (D,E) or in 60 mm culture dishes (F,G) were infected with retrovirus for control empty vector (Rt-cont), inactive hSSH1L-CS mutant (Rt-hSSH1L-CS mutant), or wild-type hSSH1L (Rt-hSSH1L WT) for 24 hours. RIE1-5 cells were transiently transfected with control (Mock), (HA)₃-Akt WT (H), LIMK1, or ROCK1 (I) plasmids, 24 hours before OXO treatment. After 24 hours without or with OXO treatment, cell lysates were prepared. An equal amount of proteins was used in standard western blotting for the indicated molecules (A,B,C,F,G,H,I). Cells on coverslips were stained for actin (D,E) or immunostained for cofilin or pS³cofilin (J). Data shown are representative of three independent experiments.