Figure 1

Vascular endothelial growth factor (VEGF) stimulation results in increased expression but not activity of RhoB protein in HUVEC. (A) RhoB protein levels are increased following VEGF stimulation. Cells were serum starved overnight in MCDB 131 with 0.25% FBS. Following this, media was replaced with MCDB 131 with 0.25% FBS and 20 ng/ml VEGF, and protein lysates were collected at the times indicated. Protein lysates were subjected to western blotting and total levels of RhoB protein were assessed. β-actin was used as a total protein loading control. (B) Levels of activated RhoB do not change following VEGF stimulation. Cells were serum starved overnight in MCDB 131 with 0.5% FBS, followed by stimulation with 20 ng/ml VEGF. At various times post-VEGF stimulation, protein lysates were collected and activated RhoB was assessed using G-LISA assays as described in Materials and Methods. Bars represent the mean ± SEM for duplicate measures in each of two independently performed experiments.