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. Author manuscript; available in PMC: 2010 Dec 11.
Published in final edited form as: Biochem Biophys Res Commun. 2009 Sep 30;390(2):252–257. doi: 10.1016/j.bbrc.2009.09.100

Figure 1.

Figure 1

Knockdown of p110α and p110β stimulate AMPK and mTOR/p70S6K phosphorylation. (A) Efficacy of RNAi-mediated knockdown of p110α and p110β was determined by real-time PCR using B2M as a reference gene. Cells were reverse-transfected with 10nM negative control siRNA (si-Con) or 5nM siRNA against p110α (si-p110α) plus 5nM si-Con, 5nM p110β (si-p110β) plus 5nM si-Con, or simultaneous transfection (co-transfection) with 5nM p110α and 5nM p110β (si-p110α \ β). Cells were maintained until harvest as described in Materials and Methods. Values are expressed relative to si-Con which was arbitrarily set at 1.0 for p110α and p110β. Columns are averages of independent duplicate determinations and error bars indicate S.E.M. (B) Cells were reverse-transfected and maintained as described in (A). Forty-eight hours after transfection, cells were harvested (left column) or treated with 125ng/ml IGF-I for 30-minutes and then harvested for protein lysates (right column). Western blotting was performed using antibodies indicated to the right of the respective images. Blots are representative of two independent experiments.