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NanoBiT assays were carried out as previously described (28): 24 hours after transfection, cells were incubated in 3 mM glucose for a minimum of 2 hours, detached and resuspended in NanoGlo Live cell substrate (N2012, Promega), 1:20 dilution in HBSS, and seeded onto an opaque white 96-well half-area plate

Cell culture The RAW264.7 cells, obtained from the Cell Bank of Type Culture Collection of Chinese Academy of Sciences (Shanghai, China), were cultured in DMEM (Corning, USA) with 10% fetal bovine serum (FBS

Regenerons Phase 2 COURAGE trial echoed these results in humans: combination therapy significantly reduced fat while preserving up to 81% of lean mass loss compared to semaglutide alone

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