Activation of AMPK by APC is mediated via the Ca2+-CaMKKβ pathway. (A) Isolated cardiomyocytes were treated with APC derivatives or saline. The rise in intracellular Ca2+ was recorded in response to electrical stimulus. Values are means ± S.E., n = 60–90 cells per group, *p< 0.05 vs. control. (B) Immunoblotting of CaMKKβ in heart and cardiomyocytes after immunoprecipitation. (C) Densitometric scanning of immunoprecipitated CaMKKβ from saline or APC-treated heart tissues following incubation with immunoprecipitated AMPK from basal saline-treated hearts with or without Ca2+ and calmodulin. (D) The phosphorylation of AMPK in mouse hearts by APC with or without pre-treatment with CaMKKβ inhibitor, STO-609. Values are means ± S.E., n=4. *p<0.05 vs. control, †p<0.05 vs. APC alone.