AMPK antagonizes DC activation. (A) Extracts of DCs stimulated with LPS for the times shown were probed by Western blotting with antibodies specific for phosphorylated Thr-172 AMPKα or for total AMPKα. (B) LPS-induced maturation, measured as p40 production and CD86 expression, as measured by flow cytometry in DCs transduced with AMPK shRNAs or control Luc shRNAs. (-) denotes resting DCs; numbers indicate percentage of CD11c+ cells within gates. (C) p40 production by resting DCs (-) and DCs stimulated with LPS or LPS plus AICAR. Cytokine production was determined by enzyme-linked immunoabsorbent assay 18 hours after activation. (D) Glucose consumption (measured as glucose remaining in culture medium after 24 hours of culture, normalized per 106 cells) by DCs cultured without or with LPS in the absence (-) or presence of 2-DG or AICAR. Bars represent mean values ± SDs from 3 independent replicates. All data are representative of 2 or more independent experiments.
