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. 2011 Jun 22;6(6):e21187. doi: 10.1371/journal.pone.0021187

Figure 3. Leucine supplementation rescues insulin signaling in muscle, liver and fat and stimulates p70S6K and AMPK phosphorylation.

Figure 3

After 8 weeks on the different diets, mice were injected i.v. with insulin (5 U per mouse) or saline, and tissues were harvested 5 minutes later. Tissue protein lysates (20 µg) were run on SDS-PAGE and subjected to western blot using antibodies directed against phosphorylated or total p70S6K, IRS-1 and AKT. Insulin receptor was immunoprecipitated with anti-IR antibody and blotted for phosphotyrosine 4G10 or insulin receptor (ß-subunit). Panels A and B show the muscle data; panels C and D, the liver data; and panels E and F, the fat data. IRS-1 phosphorylated on Ser 307 was also assessed in fat lysates (panel F), and AMPK phosphorylated on T172 was assesed in muscle lysates (panel B) by western blotting. 5 animals per group were used, the experiments and the blots were repeated 2 times.