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Cell culture INS-1 832/3 cells, a rat insulinoma subline with preserved incretin responses (64), and derivative INS-1 832/3 cells with endogenous rat GLP-1R deleted by CRISPR/Cas9 [INS-1 832/3 GLP-1R KO (27)], were grown in RPMI-1640 medium with 2 mM L-glutamine and 11 mM D-glucose, supplemented with 10% FBS, 1% p/s, 10 mM HEPES solution (H3537, Sigma-Aldrich), 1 mM sodium pyruvate (11360070, Thermo Fisher Scientific) and 0.05 mM -mercaptoethanol (M3148, Sigma Aldrich) in a humidified incubator at 37C and 5% CO 2

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