Figure 6

Gc effects on Rac1 and RhoA activity and expression. To understand effects of Gc exposure on the activity of two key regulators of cell motility, Rac1 and RhoA, pull-down assays using beads specific for the active form of these proteins was performed. This allowed quantification of the active protein: total protein. (a) Demonstrates Rac1 activity at 3 hours and 24 hours following Gc exposure. (b) Demonstrates RhoA activity at 3 hours and 24 hours following Gc exposure. Results were analysed using the nonparametric Mann-Whitney U test: vehicle-control treated cells were compared with Gc-treated cells at the 3 hour time point, and separate vehicle-control treated cells were compared with Gc-treated cells at the 24 hour time point for each experiment. The experiment was performed at least six times. N.S. = not significant. Western blot (c) and quantification (d) of total Rac1 expression following Gc exposure (n = 3), and western blot (e) and quantification (f) of RhoA expression (n = 2), showed no significant change in expression following analysis using one way ANOVA and Dunnett’s multiple comparisons test for any time point measured. Error bars represent standard error of the mean. Full length western blots are provided in Supplementary Figure S8.