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Fig. 7. The interaction of Dock180 with Crk is not required for the RhoG-induced promotion of cell migration. Motility of HEK 293T cells transiently transfected with the indicated plasmids was evaluated by Transwell migration assays. Cells were plated in the upper chamber of the filters that had been coated with fibronectin on the underside. At 6 hours after plating, cells that had migrated to the underside of the filters were fixed, and GFP-positive cells were counted. Relative cell migration was determined by the number of the GFP-positive cells that had migrated to the underside of the filter normalized to the total number of the GFP-positive cells adhering to fibronectin, and the value from cells transfected with GFP alone was arbitrarily set at 100%. (A,B) Expression of Myc-tagged RhoG, Flag-tagged Dock180 and CrkII was determined by immunoblotting with antibodies against Myc, Flag and Crk, respectively. For each experiment, the number of migrated cells was counted from the images of nine randomly selected fields on the underside of the filter. About 100 cells were assessed in one experiment, and data are the means ± s.e.m. of three independent experiments.
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