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Fig. 6. mSTI1-EGFP and its modified derivatives can interact with mSTI1, Hsp70 and Hsp90. Cells were transfected with pB-mSTI1-EGFP or derivative plasmids pB-mSTI1(S189A)-EGFP, pB-mSTI1(T198A)-EGFP, pB-mSTI1(S189E)-EGFP, pB-mSTI1(T198E)-EGFP or pB-EGFP. EGFP chimeric proteins were immunoprecipitated from transfected and control untransfected cell lysates with antibodies against GFP (anti-GFP) or non-immune IgG (IgG) and subjected to western blotting, as indicated. The mSTI1-EGFP and mSTI1 proteins were detected with a polyclonal antibody raised against GST-mSTI1 (Lässle et al., 1997). Hsp90 was detected with monoclonal antibody sc31119 and Hsc70 and Hsp70 were detected with monoclonal antibody BRM-22. The mSTI1, Hsp90 and Hsp70 proteins all specifically immunoprecipitated with the mSTI1-EGFP chimeric proteins.





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