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These observations were supported by our mutagenesis studies, where mutations R299 ECL2 A in GLP-1R, R289 ECL2 A in GIPR and I194 2.67b K in GCGR caused variable decreases in retatrutide potency (by 3.7-fold, 4.6-fold and 36.0-fold, respectively), while S297 ECL2 A in GCGR slightly increased retatrutide potency

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