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Consistently, alanine mutations at E 6.53b decreased retatrutide-induced cAMP signaling potency by 3.6-fold for GLP-1R, 1.6-fold for GIPR and 8.3-fold for GCGR, while at E/D 7.42b , potency decreased by 71.6-fold for GIPR and 5.8-fold for GCGR (Fig