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. Author manuscript; available in PMC: 2010 Jul 14.
Published in final edited form as: Oncogene. 2009 Oct 5;29(2):263–272. doi: 10.1038/onc.2009.319

Figure 1. v-Src phosphorylation of cofilin at Y68.

Figure 1

(A) 293T cells were co-transfected with His- and Myc-tagged cofilin or the mutants and HA-tagged v-Src or vector control, as indicated. Lysates were precipitated with Ni-beads, followed by western blotting with anti-phosphotyrosine, 4G10 (top) or anti-Myc (middle) antibody. Whole cell lysates (WCL) were also immunoblotted with anti-HA (bottom). (B) Recombinant GST-cofilin or GST-Y68F purified from bacteria were incubated in a kinase buffer (ATP is omitted in lane 2, as indicated by labels above the upper panel) with HA-v-Src or HA-FAK immunoprecipitated from 293T cells transfected with corresponding expression vectors, as indicated. In vitro kinase assays were performed as described in the Experimental Procedures and the reaction mixtures were subjected to immunoblotting with 4G10 antibody (upper) or anti-GST (lower). (C) 293T cells were co-transfected with Flag-tagged v-Src and HA-tagged cofilin or Y68F mutant or vector control as indicated. Lysates were immunoprecipitated with anti-HA antibody, followed by western blotting with anti-Flag antibody (upper). WCL were also immunoblotted with anti-Flag (lower). Molecular weight markers are indicated on the right.