Datasets Glycolysis and hexosamine biosynthesis pathways in valve cells

(1) Real-time metabolic analysis of human valve cells- It was performed in an Agilent Seahorse XFe24 metabolic analyzer (Agilent Technologies, Santa Clara, CA) using Cell Mito Stress and ATP assay kits. In total, 40,000 cells of endothelial valve cells or 30,000 cells of interstitial valve cells were seeded in a Seahorse XF24 Cell Culture Microplate and allowed to attach overnight. Then, cells were activated as indicated for 24 h. Before assay, media were replaced by Seahorse XF RPMI Medium, pH 7.4, supplemented with 2 mmol/L L-glutamine, 25 mmol/L D-glucose, and 1 mmol/L pyruvate. Cells were incubated for at least 45 minutes at 37°C in a non-CO2 incubator before the analysis. Data were analyzed with Seahorse Analytics software and normalized to the number of cells, evaluated by Hoechst staining in a Cytation 5 multimode reader (BioTek, Winooski, VT) at the end of the experiment. Data describe the indicated biological replicates performed in triplicate.