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FITC-agonist was added for 30 min at 37 C to induce internalization, then removed and washed with cold HBSS to arrest further endocytosis, and cells treated with 500 mM ice-cold sodium 2-sulfanylethanesulfonate (MesNa), a membrane-impermeable reducing agent, in alkaline TNE buffer (100 mM NaCl, 50 mM Tris-HCl, pH 8.6) to cleave BG-SS-biotin specifically from cell surface receptors

Month 3: Space out injections

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The usual mechanistic explanation is that the GHRH arm drives somatotroph cAMP signalling while the GHRP arm additionally attenuates somatostatin tone, so the two are acting on different limbs of the same axis rather than competing for one receptor

Why somewhere different each time