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  • 5637 and T24 BC cells, untreated or treated for different times with sorafenib (20 μM), vehicle or pretreated for 30 min with sodium orthovanadate (0.25 mM), before the treatment with sorafenib (20 μM) for 24 h, were lysed in lysis buffer (1M Tris pH 7.4, 1 M NaCl, 10 mM EGTA, 100 mM NaF, 100 mM Na V0, 100 mM phenylmethylsulfonyl fluoride, 2% deoxycholate, 100 mM EDTA, 10% Triton X-l00, 10% glycerol, 10% SDS, 0.1 M Na P 0) containing protease inhibitor cocktail (Sigma-Aldrich) by using a Mixer Mill MM300 (Qiagen, Hilden, Germany).
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