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. 2009 May 8;284(19):13013–13022. doi: 10.1074/jbc.M807653200

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Copyright © 2009, The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 6. — F411Y/S438P mutant forms γ-secretase complexes in yeast. A, microsomes from yeast transformants with PS1 wild type, NCT, Aph-1-HA, and FLAG-Pen2 were solubilized with buffer containing 1% CHAPSO. γ-Secretase complex was purified from the extracts using the antibody against PS1-loop region (G1L3) or rabbit preimmune serum, as indicated. Immunoprecipitates and the input fraction were analyzed by immunoblotting using specific antibodies. The input represents 25% of the microsomal extract. B, microsomes were prepared from yeast transformants expressing PS1 wild type or F411Y/S438P mutant, NCT, Aph-1-HA, and FLAG-Pen2 as indicated. IP, immunoprecipitation. γ-Secretase complexes were immunopurified by G1L3 antibody and analyzed by immunoblotting as A.