Fig. 3. Adenosine promotes phosphorylation of the AMPK downstream target acetyl-CoA carboxylase (ACC) in IEC-6 cells. (A) Western blot analysis of phosphorylated ACC (P-ACC) was performed after treatment for 10 minutes and 60 minutes with 10 µM adenosine (Ado) and 500 µM AICAR (Aicar), respectively; Ctrl, control. A representative immunoblot is shown. The densitometric analysis corresponds to the mean ± s.e.m. of three independent experiments. The statistical significance was assessed by Student's t-test: *P<0.05; ***P<0.001. (B) Cells were infected with either Ad.Null or Ad.1DN adenoviral vectors (30 pfu/cells). At 24 hours post-infection, cells were incubated for 10 minutes either in the presence (Ado) or the absence (C) of 10 µM adenosine. Total AMPK activity was measured in cell lysates, without prior immunoprecipitation, using the AMARA peptide assay and ACC regulation was monitored by western blot under the same conditions. As a control for infection efficiency, total 1AMPK subunit protein was measured by western blot in both Ad.Null- and Ad.1DN-infected cultures.