Figure 5

Inhibition of vitronectin-directed migration of the glioma cells from 12V-Ha-Ras transgenic mice following the administration of SF1126 and its effect on the cortical distribution of filamentous actin. A. Effect of SF1126 (50 mM) or LY294002 (50 mM) on vitronectin-directed migration of derivative glioma cells from 12V-Ha-Ras transgenic mice in transwell chambers. Bars represent the mean ± SD of number of cells migrated on vitronectin (in triplicates) in lower bottom panel. * P< 0.05. B. Effect of pre-pulse of RGDS peptide to the treatment of SF1126 and LY294002 on vitronectindirected migration of derivative glioma cells from 12V-Ha-Ras transgenic mice in scratch assays. Derivative glioma cells were allowed to migrate on vitronectin coated 24 well plates for 24 hours in presence or absence of either SF1126 (50 3 mM) or LY294002 (50 mM) following scratch on the confluent monolayer of the cells. In a separate experiment, cells were pulsed with RGDS peptide (50 mM) for 30 minutes before the administration of either SF1126 (50 mM) or LY294002 (50 mM). C. Effect of SF1126 (50 mM) on the cortical distribution of polymerized filamentous actin in the derivative glioma cells from 12V-Ha-Ras transgenic mice. D. Time-lapse confocal images of the effect of SF1126 on the vitronectindirected migration of the derivative glioma cells from 12V-Ha-Ras transgenic mice in scratch-wound assay. Vehicletreated (vehicle) and SF1126 treated (50 mM) cells were imaged separately for 8 hours (t 0 hour, t 4.2 hours, t 8.3 hours). Scale bar = 20 mm.The trajectory of the movement of the cells is characterized by two quantitative motility descriptors, average velocity (Upper bar diagram) and MRDO (maximum relative distance from the origin; Lower bar diagram) as shown in the bar diagrams. Bars represent Mean ± S.D. of the average velocity (mM/minute) and MRDO (mM) of the cells in presence of SF1126. *P<0.0025.