In vivo 2-DG uptake

Tissue glucose uptake was examined by measuring the uptake of [³H]-2-DG during an intraperitoneal GTT as described previously^46,47, with minor modifications. Mice that had been denied access to food overnight received oral administration of 30 mg kg⁻¹ of DS20060511 or vehicle 15 min prior to the glucose load. The mice then received intraperitoneal glucose administration (1 g kg⁻¹ containing 100 μCi kg⁻¹ [³H]-2-DG as tracer), followed by quick removal of the tissues 60 min later. Tissue samples were homogenized in 0.5% perchloric acid and centrifuged, and the supernatants were neutralized with KOH. One aliquot of the supernatants was used for measuring the total radioactivity ([³H]-2-DG and [³H]-2-DG 6-phosphate ([³H]-2-DGP)). A second aliquot of the supernatants was treated with 1 N Ba(OH)₂ and 1 N ZnSO₄ to remove [³H]-2-DGP, and the [³H]-2-DG count was measured. 2-DG uptake into the tissue, which is rapidly metabolized to 2-DGP in the tissue, was estimated by subtracting the count of [³H]-2-DG from the total count. ³H-specific activities were counted using a liquid scintillation counter.