Figure 5.

Increased basal phosphorylated ERK1/2 stimulated by Vav1Y3F is blocked by mAb225 pretreatment of the cells. MCF-10A cells expressing GFP or Vav1Y3F were starved in assay media containing hybridoma media as a control (C) or mAb225 overnight. The cells were then left unstimulated or stimulated with 5 ng/ml EGF for 5 minutes. Cells were lysed and PBD pulldown assays were performed. WCL samples were also blotted for levels of total Rac1, Vav1Y3FGFP, phosphorylated Pak, total Pak, phosphorylated ERK1/2, total ERK2, phosphorylated Akt, and total Akt as indicated. Data are representative of 3 independent experiments.