Figure 6.
Expression of PAK5 does not potentiate EGF-stimulated ERK/MAPK pathway activation in HEK293 cells. HEK293 cells were transfected with Flag tagged wt Raf-1, minus or plus Myc-tagged wt PAK5 or Myc tagged constitutively active PAK5. After serum starvation the cells were stimulated with EGF for 5 minutes (50 ng/ml). The cells were then lysed and separated into cytosolic and mitochondrial fractions. Equal amounts of protein from each fraction were separated by SDS-PAGE, transferred to PVDF membrane. The localization and phosphorylation of Raf-1 and ERK1/2 and PAK5 proteins were determined by Western blotting. Shown is a representative experiment from three independent experiments.