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Immunoprecipitation Five hundred g NEM-treated cell lysate prepared as described above was diluted with ice-cold KI buffer [2% (w/v) Triton X-100, 50 mM Tris-HCl (pH 8.0), 150 mM NaCl, 1 mM EDTA] and centrifuged at 15,000 X g for 10 min at 4C to obtain NEM-treated cleared cell lysate