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Immunoblotting and immunoprecipitation assays The cultured cells were rinsed once with ice-cold phosphate-buffered saline (PBS) and lysed in lysis buffer containing 20 mM HEPES (pH 7.0), 1% Triton X-100, 150 mM NaCl, 10% glycerol, 1 mM EDTA, 2 mM EGTA, 1 mM DTT, 5 mM Na 3 VO 4 , 5 mM NaF, 1 mM AEBSF, aprotinin (5 μg/ml), and leupeptin (5 μg/ml)

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Is there overlap you could cut

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