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To conjugate affinity ligands, agarose beads are first activated by using chemistry to add a reactive moiety onto agarose

As appropriate to individual experiments, a phosphatidylinositol 3-kinase (PI3K) inhibitor, LY294002, and/or a nuclear factor-B (NF-B) inhibitor, BAY11-7082, were added 1 h prior to IGF-1 treatment

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Sleep improves in days, body composition takes 6-8 weeks