The activation of MAP and PI-3 kinase pathways and the effect of GIG47 knockdown on the cell invasion. (A) Cell lysates were prepared and quantified for protein content. A total of 100 μg protein was resolved on 10% SDS-PAGE followed by immunoblot analysis with specific antibodies against total or phosphorylated forms of ERK and AKT. Immunoblot for β-actin was done as a loading control. (B) GIG47 mRNA levels were measured by real-time PCR. (C) Invasion analysis of MDA-MB-231 cells transfected with GIG47 siRNA and control siRNA was performed in serum-rich medium (50% FBS). Experiments were repeated at least three times.