NCAM induces cell migration via FGFR1, Src, and Erk1/2, whereas FGF promotes cell proliferation. (A) Monolayers of HeLa cells were scratch wounded as described in Materials and methods and left untreated (control) or stimulated with FGF-2, FGL, or NCAM-Fc. Time-lapse microscopy was performed as described in Materials and methods. Images were taken from Videos 1–4. Colored lines show five representative tracks of single cells. Plots in the right column show the trajectories of 45 individually tracked cells over the 24-h period (see Materials and methods). Values are expressed in micrometers. Bar, 30 µm. (B) Mean distance covered (top) and velocity (bottom) of cells stimulated with the different ligands. Data represent the mean ± SEM from 45 individually tracked cells from three independent experiments. (C) HeLa cells stimulated with FGF-2, FGL, NCAM-Fc, ΔFN2-Fc, or Fc were subjected to migration assays in modified Boyden chambers (see Materials and methods). FGL stimulation was also performed in the presence of PD173074, PP1, or PD98059. (D) HeLa cells transfected with an empty vector (mock) or with Myc-tagged dn-FGFR1 (transfection efficiency was nearly 100%; not depicted) were stimulated with either FGF-2 or FGL and subjected to migration assay as for C. (E) HeLa cells were subjected to cell proliferation assay in the presence of FGF-2 or FGL as described in Materials and methods. (C–E) Data represent the mean ± SEM from at least three independent experiments. *, P < 0.005 relative to untreated cells.