Fig. 8. RhoG in Rac-independent migration and cell spreading. Two clones (3 and 6) of `floxed' Rac1 MEFs in which RhoG was stably suppressed and a control (pSuper) clone were established. (A) Western blots. Upper panel: RhoG; lower panel: Rac1. (B) Stable clones were infected with GFP-Cre adenovirus (CRE) or control adenovirus (GFP). At 6-8 days after the infection, random cell migration was assayed by time-lapse imaging. Values are means ± s.e.m.; n>20 cells from three separate experiments for each condition. ^*P<0.005, ^**P<0.0005 (Student's t-test). (C) Re-expression of human RhoG in clone 3. At 5 days after infection with GFP-Cre adenovirus, cells were transfected with human GFP-RhoG (hRhoG) or control vector (RFP). Migration was assayed as before. Cells expressing GFP-RhoG were identified based on strong perinuclear fluorescence. Values are means ± s.e.m.; n>15 cells from three separate experiments. ^*P<0.005. (D) Cells prepared as in A were plated on fibronectin and spreading was analyzed as described in the Materials and Methods. Values are means ± s.e.m.; n>20 cells normalized to the size of unspread cells (5 minutes after adhesion).