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. 2012 Dec 17;209(13):2441–2453. doi: 10.1084/jem.20112607

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© 2012 Finlay et al.

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Figure 6. — mTORC1 and HIF1 do not regulate Akt activity or Foxo phosphorylation. (A) Immunoblot analysis of phosphorylated AKT and Foxos in P14-LCMV CTLs treated with and without Akti1/2 or rapamycin for 24 h. (B) P14-LCMV CTLs were treated with and without rapamycin or Akti1/2 for 24 h and subjected to nuclear/cytoplasmic fractionation before immunoblot analysis for Foxo1 and Foxo3a expression. Purity of cytoplasmic and nuclear fractions was confirmed by IκBα and Smc1 expression. (C) Immunoblot analysis of WT or HIF1^−/− CTLs for Akt–Foxo and mTORC1 signaling. HIF1^−/− CTLs were treated with rapamycin as a negative control for mTORC1 activity. For all panels, data are representative of at least three experiments. Molecular mass is indicated in kilodaltons. Dotted lines indicate that intervening lanes have been spliced out.