Cells were gently resuspended in tubes containing 10 mL of pre-warmed RPMI 1640 media (Gibco) supplemented with 10% FBS (Biological Industries), L-glutamine (2 mM), penicillin (100 U/mL, Biological Industries), streptomycin (0.1 mg/mL, Biological Industries) and Benzonase Nuclease (1:10,000, Millipore), and centrifuged for 7 min at 400 g , RT
Additionally, reductions in plasma alanine levels were observed with administration of 1.8 mg twice weekly, further supporting glucagon receptor activity
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