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. Author manuscript; available in PMC: 2012 Nov 12.
Published in final edited form as: J Immunol. 2008 Dec 1;181(11):8088–8095. doi: 10.4049/jimmunol.181.11.8088

FIGURE 2.

FIGURE 2

mTORC2 is critical for CD40-inducible activation of Akt. A, EC were transfected with control, mTOR, rictor or raptor siRNA. Thirty-six hours following transfection, the EC were serum starved for 12 h and subsequently stimulated with sCD40L (3 μg/ml) for 30 min. EC were lysed and subjected to Western blot analysis using anti-phospho Akt (pAktS473) or anti-Akt. Western blots of siRNA transfected cells confirmed efficient knockdown of mTOR, rictor and raptor in our EC (data not shown). B, Control blot in which EC were transfected with control, mTOR, rictor, or raptor siRNA, and after 48 h, Western blot analysis was performed using anti-mTOR, anti-rictor, or anti-raptor to confirm efficiency of knockdown. C, EC were serum starved for 12 h and treated with rapamycin (10 ng/ml) for an additional 12 h before stimulation with sCD40L (3 μg/ml) for 30 min. Illustrated is a representative Western blot using anti-phospho-Akt (pAktS473) and anti-Akt. One of three experiments with similar findings.