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. 2010 Oct 15;21(20):3578–3589. doi: 10.1091/mbc.E10-03-0192

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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 3. — The role of calcineurin, IGF1R and PI3-K in the activation of Akt1 in cells subjected to mitochondrial stress. (A) Effects of CnAα mRNA knockdown on the levels of Akt1 mRNA in control, mtDNA-depleted, and reverted C2C12 cells. (B) Effects of IGF1R mRNA knockdown on the levels of Akt1 mRNA in control, mtDNA-depleted, and reverted C 2C12 cells. In both A and B, Akt1 mRNA levels were quantified using real-time PCR analysis. Mean ± SEM values were calculated from three separate estimates. (C) Immunoblot analysis of total cell lysates (50 μg each) from control, mtDNA-depleted C2C12 cells, and control as well as depleted cells expressing siRNAs to CnAα and IGF1R were developed with Akt antibody. A companion blot was probed with actin antibody to assess loading levels. (D) Effects of the PI3-K inhibitor LY294002 (50 μM, 2 h) on Akt and phospho-Ser473 Akt in control and mtDNA-depleted cells. The immunoblot represents the analysis of total lysates (50 μg protein each) developed with the indicated antibodies. A companion blot was probed with actin antibody to assess protein loading.