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. 2010 Mar 18;59(6):1338–1348. doi: 10.2337/db09-1324

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© 2010 by the American Diabetes Association.

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FIG. 6. — Chronic rapamycin treatment improves insulin signaling in liver and muscle. Rats were treated with rapamycin as described in the legend to Fig. 1. A: Representative Western blots of liver lysates are shown for phosphorylated S6 (Ser240/244) and phospho-tyrosine on IRS-1 or IRS-2 immunoprecipitates (1 mg) (two representative animals of six). The graphs depict densitometric analysis of normalization of phospho-tyrosine/IRS protein. B: Representative Western blots of phosphorylated IRS-1 (Ser1101 and Ser636/639) and total proteins (two representative animals of six are shown) in liver lysates. The graphs depict densitometric analysis of normalization of phospho-IRS-1/IRS-1 protein. C: Representative Western blots of total IRS-1 and IRS-2 protein in liver lysates. The graphs depict densitometric analysis of normalization of total IRS/Actin. D: Equal amounts of liver protein (1 mg) were immunoprecipitated with IRS-1 and IRS-2 antibodies. Immunoprecipitates were analyzed for PI 3-kinase activity. The graphs depict densitometric analysis of IRS-1– and IRS-2–associated PI 3-kinase activity. E: Representative Western blots of liver lysates are shown for phosphorylated Akt (Ser473 and Thr308) and total proteins (two representative animals of six). The graphs depict densitometric analysis of normalization of phospho-Akt/Akt protein. F: Liver proteins (500 μg) were immunoprecipitated with total Akt antibody. Immunoprecipitates were analyzed for Akt activity. The graphs depict densitometric analysis of total Akt activity. G: Representative Western blots of muscle lysates are shown for phospho-tyrosine on IRS-1 immunoprecipitates (1 mg). The graphs depict densitometric analysis of IRS-1 phospho-tyrosine corrected for IRS-1 protein content. n = 6 animals. H: Muscle proteins (500 μg) were immunoprecipitated with total Akt antibody. Immunoprecipitates were analyzed for Akt activity and the graph depicts the densitometric analysis of several independent determinations. n = 6 for each group. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001.