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Fig. 2. Verification of paxillin {delta} as an internal translation product and its localization to focal adhesions. (A) CHO.K1 cells were transfected with either pcDNA3.1 vector only, wild-type avian paxillin (wt pax), full-length avian paxillin with internal Kozak mutation (FL pax KM) or avian paxillin {delta} (pax {delta}) (amino acid 133 to end). Immunoprecipitation with avian-specific paxillin polyclonal antiserum, Pax1, was performed on detergent-soluble lysates (DSL). Lysates and immunoprecipitates were resolved by 7.5% SDS-PAGE and transferred to nitrocellulose for western blot analysis. Paxillin monoclonal antibody PXC10 was utilized for western blotting. (B) Wild-type paxillin (a,d), full-length paxillin KM (b,e) and paxillin {delta} (c,f) were transiently transfected into CHO.K1 cells followed by respreading of the cells on fibronectin-coated coverslips. Cells were processed for indirect immunofluorescence microscopy utilizing the avian-specific Pax1 antibody (a,b,c) and double-labeled with an antibody to vinculin (d,e,f). Bar, 5 µm.





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