Skip to main content
. Author manuscript; available in PMC: 2014 May 9.
Published in final edited form as: Mol Cell. 2013 Apr 18;50(3):356–367. doi: 10.1016/j.molcel.2013.03.015

Figure 3. Knockdown of Akt Reduces Ago2-mediated mRNA Repression.

Figure 3

(A) Ago2 S387 phosphorylation positively affects CXCR4 siRNA-mediated repression. HeLa cells harboring endogenous Ago2 knockdown and expressing ectopic Ago2 mutants were assayed for CXCR4 siRNA targeting to the FL6X mRNA repression reporter. (B) Knockdown of Akt3 significantly disrupts miR-21-mediated repression in HeLa-D8 cells. (C) Phospho-dominant Ago2-S387E mutant partially rescues the Akt knockdown phenotype. HeLa-D8 cells were co-transfected with siRNAs targeting Akt3 and Ago2 and Ago2 constructs. (D) Knockdown of Akt3 increases PTEN protein but not pten mRNA levels in HeLa cells. Bars indicate pten mRNA levels (normalized to GAPDH) and triangles represent pten protein levels relative to the NT control sample. (E) Knockdown of Akt3 up-regulates translation of miR-21 targets PTEN and PDCD4 in HEK293T cells measured by 35S methionine incorporation. Relative amounts of PTEN and PDCD4 proteins are indicated using siNT as a reference for input and IP. Error bars indicate SD. *P ≤ .05, **P ≤ .01, ***P ≤ .001. See also Figure S3.