Figure 6

Anti-L1 antibodies and RGD peptide inhibit glioma cell motility. (A) Anti-L1 antibody ASCS4 was added to 9L/LacZ cultures and assessed by the Super Scratch assay for effects on cell motility. The time-lapse graph shows the average velocities and best-fit curves of the control and antibody treated 9L/Lacz cells over a 20 hour time period (15 cells each condition). The antibody ASCS4 consistently lowered the velocities of migrating cells throughout the course of the experiment. (B) The overall average velocities were calculated using all individual cell velocities collected during the course of the experiment and graphed as single values for the two populations of cells. Control cell velocity was 0.073 ± 0.002 μm/min. Velocity of the ASCS4 treated cells was reduced 34% compared to untreated controls at 0.048 ± 0.001 μm/min (p < 0.001). Bars are s.e.m. (C) Anti-L1 antibody EZ1 or the RGD-containing peptide antigen were added to cell cultures as described and assessed by the Super Scratch assay for effects on 9L/LacZ cell motility. The time lapse graph shows the average velocities and best-fit curves of the control, antibody treated, and peptide treated 9L/Lacz cells over a 20 hour time period (2 areas of interest and 40 cells each condition). The antibody EZ1 and peptide consistently lowered the velocities of migrating cells throughout the course of the experiment, with the antibody having a greater effect on reducing migration velocity. (D) The overall average velocity for the three populations of cells from (C) is shown. Control cell velocity was 0.214 ± 0.003 μm/min. Velocity of the EZ1 treated cells was reduced 32% (0.146 ± 0.003 μm/min) and velocity of peptide treated cells was reduced by 19% (0.174 ± 0.003 μm/min) compared to untreated controls (p < 0.001 for each comparison). Bars are s.e.m. Shown are data from one representative experiment. The experiments were repeated twice with similar results.