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For flow cytometry experiments, cells were prepared as indicated in the flow cytometry section and incubated for 30 min at 37 C with Calcein-AM (10 M) or Rhonox (5 M) before being washed twice with PBS and suspended in PBS containing 0.5% serum prior to being analyzed

The compound shows high enzymatic selectivity, preserves lean mass, and engages longevity-related pathways (e.g., SIRT1), but remains a nonFDA-approved research substance that is WADA-banned and should be used under medical guidance

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Furthermore, PSAs confirmed that subcutaneous tirzepatide remained cost-effective compared with oral semaglutide

Similar results were found at mRNA expression level with downregulation of ZO-1 and occludin before the onset of arthritis (Supplementary Fig