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. 2012 Jul 10;287(35):29406–29416. doi: 10.1074/jbc.M112.348946

FIGURE 2.

FIGURE 2.

CCL5 activates the energy-sensing kinase AMPK and the downstream substrate GSK-3β resulting in an increased intracellular ATP levels. A, activated PB T cells were either left untreated or treated with 10 nm CCL5 for the indicated times. Cells were harvested and protein lysates resolved by SDS-PAGE and immunoblotted with anti-phospho-AMPKα (Thr-172) or anti-phospho-GSK-3β (Ser-9) antibodies. Membranes were stripped and reprobed for loading. Relative phosphorylation is shown as signal intensity over loading control. Data are representative of two independent experiments. B, activated PB T cells were either treated with dimethyl sulfoxide or 10 μm compound C for 1 h before treatment with 10 nm CCL5 for the indicated times. Intracellular ATP was measured using a bioluminescent assay. Data are representative of two independent experiments. *, p < 0.01; **, p < 0.05. C, activated PB T-cells were pretreated with dimethyl sulfoxide (DMSO; carrier control), oligomycin (1 μm), or 2-deoxy-glucose (10 mm) for 30 min and then stimulated with IL-2 (20 ng/ml) or CCL5 (10 nm) for 30 min. Intracellular ATP levels were then measured using a bioluminescent assay. Data are representative of two independent experiments. *, p < 0.01.