AMPK is activated and phosphorylated in response to AICA riboside and phenformin in LKB1+/+ but not in LKB1-/- MEF cells. Cells immortalized from control embryos LKB1+/+ and LKB1-/- knockout embryos , were incubated with AICA riboside (2 mM) or phenformin (10 mM) for 1 hour. Lysates were prepared and AMPK activity (expressed as units per mg total lysate protein) determined in immunoprecipitates made using a mixture of anti-AMPKα1 and anti-AMPKα2 antibodies. Lysates were also analyzed by SDS gel electrophoresis and blots probed using anti-LKB1, anti-AMPKα1/β2, and anti-pT172 antibodies.
Hawley et al. Journal of Biology 2003 2:28 doi:10.1186/1475-4924-2-28