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. 2012 May 2;287(28):23489–23501. doi: 10.1074/jbc.M112.365874

FIGURE 5.

FIGURE 5.

AMPK activation induces dissociation of SIRT1 from DBC1. A and B, SIRT1-DBC1 interaction was evaluated by co-immunoprecipitation in A549 cells. A, cells were treated with resveratrol (100 μm), oligomycin (5 μm), or A769662 (100 μm) for 2 h before performing immunoprecipitation (IP) for DBC1. Proteins were immunoblotted with anti-SIRT1, anti-DBC1, anti-phosphorylated AMPK (P-AMPK) (Thr-172), and anti-AMPK antibodies. B, cells were treated with resveratrol (RSV; 100 μm) or resveratrol + compound C (CC; 10 μm) for 2 h before performing immunoprecipitation. Immunoprecipitates were immunoblotted with anti-SIRT1 and anti-DBC1 antibodies. The graph shows the average of three independent experiments. * Denotes difference to control, and ** denotes difference to RSV+CC. Error bars represent S.D. C, SIRT1-DBC1 interaction was evaluated by co-immunoprecipitation in 293T cells after transfection of FLAG-SIRT1, Myc-DBC1, and CA AMPα or dominant-negative AMPKα (DN). Immunoprecipitates were immunoblotted with anti-FLAG and anti-Myc antibodies.