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Description

For co-immunoprecipitation assays, human IR, IR Y972F and IR L973F cDNA were cloned into 3 Flag-CMV-14 mammalian expression vector (Sigma)

Culture media consisted of 10 g.L -1 glucose, 1 g.L -1 yeast extract, 5 g.L -1 (NH 4 ) 2 SO 4 , 2 g.L -1 KH 2 PO 4 , 0.5 g.L -1 MgSO 4 .7H 2 O, 0.1 g.L -1 CaCl 2 .2H 2 O, 0.02 g.L -1 FeSO 4 .7H 2 O, 0.01 g.L -1 ZnSO 4 .7H 2 O, 0.005 g.L -1 CuSO 4 .5H 2 O, 0.001 g.L -1 MnCl 2 .4H 2 O, 2.5 mL 70% H 2 SO 4 and 0.5% (v/v) Antifoam 204 (Sigma Aldrich, St