Anti Rhodamine  

SKAP2 directly inhibits actin polymerization induced by the WAVE2-cortactin complex.？A, top,？diagram of the actin polymerization assay. Rhodamine-conjugated monomeric G-actin, WAVE2, and GST-SKAP2 were added to cortactin-coated protein G beads. HeLa cell lysates were also added to supply the Arp2/3 protein complex. After incubation, protein G beads were separated from the reaction mixture, washed, and subjected to immunoblotting (IB).？Bottom,？polymerized actin, which was produced in the complex of cortactin and WAVE2, was detected using anti-rhodamine and anti-actin antibodies. WAVE2, SKAP2, and cortactin were detected using anti-WAVE2, anti-SKAP2, and anti-EGFP antibodies, respectively.？Sup, reaction mixture;？beads, beads fraction after incubation.？B, top,？diagram of the actin polymerization assay using EGFP-WAVE2-coated beads.？Bottom,？polymerized actin was analyzed as？A. A？and？B,？signal intensities of rhodamine-actin in the beads fraction were quantified. Signal intensities of lanes with WAVE2 without SKAP2 were defined as 1. The results from the three independent experiments are shown as the means ± S.D. Fig 7.PMID 23161539.