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. 2006 Feb 27;172(5):771–782. doi: 10.1083/jcb.200512115

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Copyright © 2006, The Rockefeller University Press

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Figure 8. — Localization of proteins in wild-type and zyxin-null fibroblasts demonstrate the importance of zyxin for proper localization of Mena and VASP. Protein localization in wild-type (+/+) and zyxin-null (−/−) cells was evaluated by indirect immunofluorescence microscopy using antibodies specific for zyxin (A and B; rabbit polyclonal antiserum B71), vinculin (C and D), Mena (E and F), and VASP (G and H). Wild-type cells display colocalized Mena (I) and vinculin (J) at focal adhesions. Mena failed to localize normally to zyxin-null focal adhesions (K) although vinculin persisted (L). Expression of GFP-zyxin in zyxin−/− cells restored the localization of Mena to focal adhesions (M), colocalizing with vinculin (N). To show the enhanced LPP focal adhesion staining (arrows) in fibroblasts lacking zyxin, a mixed population of wild-type (+/+) and zyxin-null (−/−) cells was stained for LPP (O) and for zyxin (P; mouse monoclonal antibody 1D1).