Figure 7
HGF-FN and HGF-VN induced HMVEC migration is Ras dependent. Panel A-GTP-Ras pull down assay showing the affects of FPT III (100 μM) and GGTI (2 μM) inhibitors upon GTP-Ras levels. HMVEC suspensions (3 × 106/ml) were pre-treated with FPT III and GGTI inhibitors for 45 min at room temperature prior to stimulation with HGF-FN complexes for 60 min at room temperature. Samples were analysed as mention in the legend to Fig 2. Panel B-Effect of Ras inhibition upon cell migration. Calcein M loaded HMVEC suspensions (1 × 105/ml) were pre-treated with the inhibitors as mentioned above prior to application to the top chamber of the transwell filter. Cells were stimulated with HGF plus ECM proteins (placed in the bottom chamber) as shown. Cell migration was measured at 3 hours post stimulation using a fluorescence plate reader. The data is shown as relative migration and is the combined data from two independent experiments with sample wells in triplicate.