Fig. 2

RA reduced endothelial glycolysis. a Extracellular acidification rate (ECAR) profile in DMSO and RA-treated hESC-ECs. The vertical lines indicate the times of addition of glucose (10 mmol/L), oligomycin (3 μmol/L), and 2-deoxy-D glucose (100 mmol/L). Quantification of glycolysis (b), glycolytic capacity (c), and glycolytic reserve (d) for the cells described in panel (a). (e) Representative flow cytometric overlay and quantification of 2-NBDG uptake in RA-treated hESC-ECs compared with control hESC-ECs, as measured by flow cytometric analysis of 2-NBDG fluorescence. f Statistical analysis of the data in panel e showing significantly decreased 2-NBDG uptake. g Representative photographs of endothelial spheroids showing reduced sprouting in 2-DG-treated hESC-ECs. Scale bar, 100 μm. h Statistical analysis of the data in panel g showing a significantly decreased total sprout length and sprout/spheroid ratio. All data are presented as mean ± SEM; Student’s t test; *p < 0.05, **p < 0.01, and ns, not significant