Fig. 6 | Nature Communications

Fig. 6

From: Endothelial adenosine A2a receptor-mediated glycolysis is essential for pathological retinal angiogenesis

Fig. 6

Glycolysis is involved in ADORA2A activation-mediated HRMEC proliferation and sprouting. ag Results of Ki-67 and BrdU staining ac, WST-1 cell proliferation assay d and EC spheroid sprouting assay eg in HRMECs. HRMECs were infected with Ad-Ctrl or Ad-A2AR adenoviruses, and then treated with adenosine in the presence or absence of two different glycolytic inhibitors (3PO, 10 μM, and 2-DG, 5 mM). n = 6. *P < 0.05; **P < 0.01; ***P < 0.001. Scale bars: 50 μm for a; Scale bars: 100 μm for e. hj HRMECs were cultured as 3D multicellular spheroids in the presence or absence of 3PO or 2DG. Representative images of spheroidal sprouting after culturing for 24 h in collagen matrix under hypoxia (0.5% O2) or normoxia (21% O2) h. Morphometric quantification of spheroid sprouting by calculating the number of sprouts per spheroid i and total sprout length j. n = 10 per group (n is number of spheroids quantified). *P < 0.05, **P < 0.01; ***P < 0.001. Scale bars: 100 μm. Data are represented as means ± s.e.m. Statistical significance was determined by one-way ANOVA followed by Bonferroni test

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