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One g of RNA was reverse transcribed into cDNA using the Transcriptor First Strand cDNA synthesis kit (Roche, Indianapolis, IN, USA) and subjected to quantitative PCR using the SYBR-Green Supermix (Roche, Indianapolis, IN, USA) with the ABI Stepone Plus real-time PCR detection system (Applied Biosystems, Foster City, CA, USA)

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Retatrutide adopts a single continuous helix that penetrates the core of the receptor transmembrane domain (TMD) via its N-terminal segment (residues 113), while its C-terminal segment (residues 1430) interacts with the N-terminal -helix of the ECD, the extracellular tip of TM1 and ECL1 (Fig