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. 2010 Sep 15;21(18):3258–3268. doi: 10.1091/mbc.E10-05-0454

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© 2010 by The American Society for Cell Biology

This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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Figure 1. — Overexpression of Rheb induces mTOR signaling through a PI3K-independent mechanism in skeletal muscle myoblasts. C2C12 myoblasts were transfected with GFP or cotransfected with a combination of GFP and GST-tagged p70^S6k (GST p70^S6k) or HA-Rheb and GST p70^S6k. At 48 h after transfection, the myoblasts were switched to serum-free media for 2 h and then incubated with 500 nM wortmannin or the solvent vehicle (DMSO) for an additional 25 min. Where indicated, the myoblasts were stimulated with 100 nM insulin during the final 10 min of this incubation period. Cell lysates were subjected to Western blot analysis with the indicated antibodies. The ratio of GST P-p70^S6K(389) to total GST p70^S6K (GST P-p70/Total GST p70) was calculated and is expressed as a percentage of the drug treatment control samples (vehicle, GFP + GST p70^S6K or wortmannin, GFP + GST p70^S6K). Values are the mean + SEM, n = 4–6/group. *, significantly different from drug treatment control.

	
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