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Glucose uptake assay

KW Kota Wakiyama
YK Yoshihiko Kitajima
TT Tomokazu Tanaka
MK Masao Kaneki
KY Kazuyoshi Yanagihara
SA Shinichi Aishima
JN Jun Nakamura
HN Hirokazu Noshiro
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Glucose uptake in cultured cells was determined using a 2-Deoxyglucose (2-DG) Uptake Measurement Kit (COSMO BIO Co. Ltd., Tokyo, Japan) as previously described16. Briefly, cells were cultured under a serum-starved condition for 6 h, followed by further culture for 18 h in regular medium supplemented with 10% FBS. The cells were then incubated for 24 h under normoxia or hypoxia. Thereafter, the cells were treated with or without 500 ng/ml insulin for 18 h. Finally, the cells were treated with 2DG for 20 min and subjected to 2DG uptake measurements according to the manufacturer’s instruction. All experiments were performed in triplicate, and mean values were calculated.

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