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For saturation binding assays, cell lysates (100 L, 25 g of protein for HEK-293 cells and 1020 g of protein for INS-1E cells, quantified by the Bradford Assay) were incubated with the indicated concentrations of [ 3 H]RX821002 (90 min, room temperature)

Similar findings have been demonstrated by agouti-related peptide through melanocortin-3 and -4 receptors (MCR3, MCR4) or by endogenous opioid peptides at - and -opioid receptors 86,87,88

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Five-year data showed sustained benefit with continued use

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