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The samples were homogenized in lysis buffer (mixture of Tris-HCl pH 8.0, 50 mmol/L of sodium chloride, 1.0% NP-40, 0.5% sodium deoxycholate, 0.1% sodium dodecyl sulfate, and protease inhibitors), and the protein concentration was determined using a bicinchoninic acid (BCA) protein assay kit (Beyotime Biotechnology Co., Ltd, China) after homogenate centrifugation