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. Author manuscript; available in PMC: 2012 Mar 1.
Published in final edited form as: Cancer Res. 2011 Jan 21;71(5):1730–1741. doi: 10.1158/0008-5472.CAN-10-1432

Figure 2. Arg is required for cortactin phosphorylation at invadopodium precursors.

Figure 2

(A) MDA-MB-231 cells stably expressing cortactin-TagRFP were treated with siRNA specific for control, Arg, Abl, or Src, plated on fibronectin/gelatin matrix and starved before EGF stimulation. Cells were fixed and labeled for phosphorylated cortactin using a phosphospecific cortactin antibody (anti-pY421) before (0min.) or after (3min.) EGF stimulation. (B) Cortactin phosphorylation was measured as the ratio of pY421 signal/cortactin-TagRFP signal at invadopodium precursors. Control siRNA: n=105(0min.)/n=127(3min.), Abl siRNA: n=110(0min.)/n=97(3min.), Arg siRNA: n=138(0min.)/n=136(3min.), Src siRNA: n=122(0min.)/n=160(3min.). Scale bars=10 μm. (C) MDA-MB-231 cells were treated with increasing concentrations of EGF for 3 minutes, lysed, immunoprecipitated with anti-cortactin antibodies followed by immunoblotting with anti-Arg antibodies. The samples were blotted with anti-cortactin as control.