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Cells were collected at indicated timepoints (0, 2, 4, 6, 12 and 24 h) and lysed in lysis buffer (20 mM Tris, pH 7.5, 150 mM NaCl and 1% Triton X-100) containing protease inhibitors on ice for 10 min, followed by centrifugation at 12,000 g for 15 min at 4 C