AICAR activates metabolic AMPK targets but largely does not affect metabolic gene expression. A, C) Immunoblotting on whole heart extracts from left ventricle heart sections after 6 d of saline or AICAR injections. B) RNA was isolated from left ventricle heart sections and qPCR was run from unstressed-saline or AICAR-injected animals after 6 d of injections. Expression is normalized to GAPDH. n = 4–6. D) NAD/NADH ratios were calculated from freshly harvested whole heart tissue from mice subjected to saline or AICAR injections for 6 d; n = 5 (WT + saline), 6 (WT + AICAR), 3 (CKO + saline, 3 were under the detection limit), and 6 (CKO + AICAR). Results are presented as the means ± sem analyzed by 2-way ANOVA with multiple comparisons and with a Tukey’s post hoc analysis. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001.