Annexin A2 is newly synthesized during the EGF stimulated migration and invasion of MDA-MB-231 cells. (A) 95 proteins in total were identified following mass spectrometry analysis and MASCOT peptide identification. Of these, 41 were found in the migration experiment only, 40 were found in the invasion experiment only and 14 were found to be common to both, listed here. (B) Protein-protein interactions between the 14 proteins found to be common to both the migration and invasion lists were assessed with String (http://string-db.org) [16] and visualized using the StringApp in Cytoscape set at 0.2 confidence level [17]. Edge line thickness denotes the confidence of association between protein nodes. (C) (i) Input: Western blot analysis of Click-It reaction lysate shows successful incorporation of the TAMRA tag into newly synthesized proteins when AHA was added. (Lane 1: Cell lysate of EGF migrating cells without AHA, Lane 2: Cell lysate of EGF migrating cells with AHA). (ii) Immunoprecipitation: Consequent TAMRA immunoprecipitation assay showed Annexin A2 pulldown following EGF induced cell migration as measured by SDS-PAGE and western blotting with anti-Annexin A2 antibody. (Lane 1: Beads only control, Lane 2: AHA+ lysate from lane 2 of (i), Lane 3 AHA- lysate from lane 1 of (i)).