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The in vitro kinase assays were conducted in a kinase reaction buffer (50 mM Tris-HCl pH 7.4, 1 mM dithiothreitol (DTT), 10 mM MgCl 2 , 0.5 M ATP) containing the IPed AMPK complex and active GSK-3 (100 ng) plus 2 g soluble various forms of GST-SIRT7 proteins