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Extracellular acidification rate (ECAR) analysis

YJ Yu Jiang
QH Qiuju Han
HZ Huajun Zhao
JZ Jian Zhang
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For the ECAR analysis, indicated THP-1-M was plated onto the Seahorse XFe24 Cell Culture Microplates (Agilent, Palo Alto, CA, USA). When cells were adhered, the medium was changed to Seahorse XF Base Medium (with 1 mM GlutaMax, pH 7.4; 102,353–100; Agilent) and incubated in a non-CO2 incubator for 1 h at 37 °C prior to the start of the assay. The assay was performed with Seahorse XF Glycolysis Stress Test Kit (103020–100; Agilent) according to the manufacturer’ s instructions. Briefly, the test consisted of four consecutive stages: basal (without drugs), glycolysis induction (10 mM glucose), maximal glycolysis induction (1 μM oligomycin), and glycolysis inhibition (50 mM 2DG). The data were collected and analysed using the XFe24 wave software (Agilent).

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