Xfe24 wave software

Seahorse extracellular flux analyzer (Seahorse Bioscience) was used to measure cellular OCR and cellular PER as the manufacturer's protocol as reported 12 (link), 28 (link). Data were analyzed by the Agilent Seahorse XFe24 Wave software. The data of OCR and glycolytic rate measurements were standardized to the cell count. Both OCR and PER were expressed as pmol/min/cells.

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).