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The gradient profile was: 3% solvent B for 5 min, increased to 20% over 4 min, further increased to 50% over 2 min, then to 95% over 1 min, followed by re-equilibration to 3% for 2 min (total run time 14 min)

Keep GHK-Cu spray refrigerated between 2-8C, protected from light, and avoid freezing

After incubation (10 min, 39C), the plates were centrifuged (200 x g, 5 min, 20C, Universal 320, Hettich, Tuttlingen, Germany) and the medium was replaced with pre-warmed XF Seahorse medium supplemented with glucose, sodium pyruvate and glutamine (all purchased from Agilent, Santa Clara, California, USA)

SIRT3-mediated SOD2 and PGC-1 contribute to chemoresistance in colorectal cancer cells

The resulting pharmacokinetic profile changes the timing and amplitude of GH and downstream IGFI signals, which can impact reproductive hormone systems associated with fertility