|The transfected cells were washed with phosphate-buffered saline and lysed in NP40 lysis buffer (20 mM Tris-HCl [pH 7.6], 150 mM NaCl, 5 mM EDTA, 1% NP-40, 0.5% sodium deoxycholate, and 0.1% sodium duodecyl sulphate) containing a complete protease inhibitor cocktail (Boehringer-Mannheim, Mannheim). The cell extracts were centrifuged at full speed in a refrigerated Eppendorff centrifuge for 10 minutes.|
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