Fig. 5

Rho activation assays. a Active endogenous RhoA levels assessed by pulldown with a GST-rhotekin Rho binding domain (GST-RBD) from HEK293T cells transfected with a gradient of Flag-p190RhoGAP-A, pG1-pG2 deletion mutant (ΔpG1–pG2), or empty Flag vector (−). A representative set of immunoblots is shown. b Scatter plot of active RhoA as a function of Flag-p190RhoGAP expression. Active RhoA is normalized to maximum levels in each experiment (empty vector control) and Flag-p190RhoGAP expression as a percentage of maximum expression within each experiment. Shown is the compilation of data from 8 independent experiments. c Scatter/bar plot of data from (b), with data points binned according to Flag-p190RhoGAP expression as indicated (1–10%, 10–25%, 25–50% and 50–100%). The mean and s.e.m. (error bars) of each group is shown. Unpaired t-tests (two-tailed) were performed in GraphPad Prism, with statistical significance indicated above pairs and p-values as follows: 1–10% bin p=0.0053; 10–25% bin p=0.0019; 25–50% bin p=0.0006; and 50–100% bin p=0.0133