Fig. 5

SphK1 promotes angiogenesis by regulating the glycolytic modulator PFKFB3. A Primary HUVECs were treated with 5 µM PF-543 for 16 h, prior to stimulation with 50 ng/ml VEGF-A for 15 min. B Following 1 h pre-treatment with 10 µM MG-132, primary HUVECs were treated with PF-543 at 5 µM for 16 h. A and B Western blotting analyses of the indicated proteins; n = 3. C and D Glycolytic rate, capacity and reserve were determined using Seahorse real-time glycolytic stress assay in PF-543-treated (C) or shRNA-mediated SphK1 knockdown (D) primary HUVECs. ECAR, extracellular acidification rate; n = 5. Data are expressed as mean ± SD. *p < 0.05; **p < 0.01; ***p < 0.001