Figure 3
Effect of S1P on cell migration and invasion.
Mitomycin C-treated colonies in serum free media were either left untreated or treated with S1P (1 and 5 Μm); (A) or with 5 μM S1P in the presence or absence of chemical inhibitors of Rac-1 (NSC23766; 5 μM) or MEK1/2 (UO126; 10 μM) (B) for 48 hours. Epithelial growth factor (EGF) was used as a positive control of cell scatter. Representative images from three independent experiments are shown. The extent of cell scattering was analysed by measuring the density of colonies of H357 and H413 cells (cells/μm2). The data represents the average of measurements taken from thirty colonies chosen at random. (C) Migration was measured using transwell assays using H357 and H400 cells in the absence or presence of 1 or 5 μM S1P. Results are expressed as percentage of migrated cells in the untreated control (=100%) ± SD. (D) Invasion assay was performed by plating H357 or H400 cells on collagen in the presence or absence of 5 μM S1P. Results represent data obtained from three independent experiments. ***ρ < 0.001, **ρ < 0.01 and *ρ < 0.05 (Dunnett’s post-hoc test or unpaired T-test).
