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Measurement of intracellular glutamine and glutamate levels

ZD Zheng-Lin Dong
XJ Xin Jiao
ZW Zeng-Guang Wang
KY Kai Yuan
YY Yi-Qi Yang
YW Yao Wang
YL Yun-Tao Li
TW Tian-Chang Wang
TK Tian-You Kan
JW Jian Wang
HT Hai-Rong Tao
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We used an intracellular glutamine assay kit (Dojindo, Tokyo, Japan) and a glutamate assay kit (Solarbio, Beijing, China) to determine the concentrations of the respective substances within the cells. In brief, after cell lysis, centrifugation was performed to collect the supernatant. Subsequently, gradient dilution was carried out using standard solutions. Then, the absorbance of the gradient-diluted standard solutions and samples was measured using an Infinite M200 Pro multimode microplate reader (Tecan Group, Ltd., Männedorf, Switzerland). A standard curve was constructed based on the absorbance values of the gradient-diluted standard solutions. The level of the corresponding substance in the samples was predicted using the standard curve. In this experiment, NP cells were respectively treated with si-NC, IL-1β + si-NC, IL-1β + mannose + si-NC, IL-1β + mannose + si-Txnip, or IL-1β + mannose + si-Txnip + si-Myc.

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