Figure 3

Representative overlay 2D gels images of MDA-MB-231 cells and p110γ KD cells. (A) Overlay gel image of the cytosolic proteome of MDA-MB-231 cells and p110γ knockdown cells without stimulation. Cytosolic fractions from triplicate samples were labeled with Cy3 or Cy5 while the pool of these six samples was labeled with Cy2 as the internal pooled standard. These labeled samples were combined and subjected to 2D gel separation, followed by scanning at 100 μm resolution using a Typhoon Trio. These data are representative of 3 gels run with similar results. (B) Cy5-labeled representative gel of the protein fractions from p110γ knockdown cells. Number-labeled rectangles indicate the protein spots with altered abundance. (C) Overlay gel image of the cytosolic proteome of MDA-MB-231 cells and p110γ knockdown cells after 5 min stimulation with IGF-1. Cytosolic fractions from triplicate samples were labeled with Cy3 or Cy5 while the pool of these six samples was labeled with Cy2 as the internal pooled standard. These labeled samples were combined and subjected to 2D gel separation, followed by scanning at 100 μm resolution using a Typhoon Trio. These data are representative of 3 gels run with similar results. The white rectangle indicates the area containing the spots 101, 102 and 106. (D) Cy5-labeled representative gel of the protein fractions from p110γ knockdown cells. Number-labeled rectangles indicate the protein spots with altered abundance.