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The fixed organoid cultures were blocked in 5% donkey serum (Millipore, Billerica, MA, USA) + 0.3% Triton-X-100/PBS, incubated overnight in 3% bovine serum albumin (Sigma-Aldrich, St Louis, MO) + PBS with primary antibodies, washed, incubated with Alexa Fluor-conjugated secondary antibodies (Invitrogen, Carlsbad, CA), washed, and stained with DAPI or mounted in VECTASHIELD H-1000

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Option Cagrilintide Evidence for the added compound Human Phase 2 data alone and Phase 3 data with semaglutide Evidence for the retatrutide combination None identified Option Tesamorelin Evidence for the added compound Randomized trials and an FDA-approved narrow indication Evidence for the retatrutide combination None identified Option CJC-1295 + ipamorelin Evidence for the added compound Small human studies showing GH or IGF-1 response Evidence for the retatrutide combination None identified Option MOTS-C Evidence for the added compound Human observational biology plus mostly preclinical treatment research Evidence for the retatrutide combination None identified This ranking describes research depth

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