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Figure 1


Fig. 1. Adenosine activates AMPK in IEC-6 cells. (A) IEC-6 cells were incubated either in the presence or absence of 10 µM adenosine (Ado) or 500 µM AICAR (Aicar) for 10 minutes and 1 hour, respectively; C, control. AMPK complexes were isolated from cell lysates by immunoprecipitation with antibodies to either anti-{alpha}1AMPK (open bars) or anti-{alpha}2AMPK antibody (solid bars). {alpha}1AMPK and {alpha}2AMPK complex activities were measured by phosphorylation of AMARA peptide. Activities shown are the mean and s.e. of five and three independent experiments for {alpha}1AMPK and {alpha}2AMPK, respectively, with each assayed in duplicate. The statistical significance of the adenosine and AICAR effects was established by Student's t-test (**P<0.01; ***P<0.001). (B) Cells were treated with either adenosine or AICAR or control (Ctrl), as detailed above. Equal amounts of protein (40 µg) were loaded to allow direct comparisons of phosphorylated AMPK (P-AMPK) levels. ß-actin was used as a loading control. A representative western blot is shown.





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