Skip to main content
. Author manuscript; available in PMC: 2013 Mar 1.
Published in final edited form as: Cancer Res. 2012 Jun 22;72(17):4394–4404. doi: 10.1158/0008-5472.CAN-12-0429

Figure 3. AMPK-α2 suppressed the activity of SIRT1.

Figure 3

A) HepG2 cells were co-transfected with increasing dosage of plasmid encoding an activated AMPK-α2 mutant (T172D) and p53. Western blotting was performed using phospho-specific (Ser20) and acetylation-specific (Lys382) anti-p53 antibodies. B) GST-AMPK catalytic domain T172D (GST-AMPK-CAD) was incubated with wild type, Ser15A and Ser20A mutants of GST-p53 in a in vitro kinase assay. The autoradiograph and the Coomassie blue staining of protein bands were shown. C) HepG2 cells were co-transfected with p53 and SIRT1 and incubated with increasing concentrations (50 μM and 100 μM) of the AMPK activator, A769662, for 4 hours. The effect of p53 acetylation was studied by Western blotting with acetylation specific anti-p53 antibody. D) HepG2 cells were co-transfected with a p53-luciferase reporter and wild-type SIRT1 with increasing amount of AMPK-α2 T172D expressing plasmids and luciferase acitivity was measured. Results are mean ± SD; *, P<0.01. Each set of samples was in triplicate and each set of experiments was repeated for 3 times.