Fig. 3 | Nature Communications

Fig. 3

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

Fig. 3

Hypoxia upregulates ADORA2A expression by activating HIF-2α in HRMECs. a Real-Time PCR analysis of mRNA expression for adenosine receptors in HRMECs. HRMECs were exposed to normoxia (21% O2) or hypoxia (0.5% O2) for 6 h. n = 4. ***P < 0.001 vs. normoxia. b, c Real-Time PCR analysis of ADORA2A mRNA expression in HRMECs. HRMECs were exposed to normoxia (21% O2) or hypoxia (0.5% O2) for indicated times b or with indicated O2 concentrations for 6 h c. n = 4. *P < 0.05; **P < 0.01, ***P < 0.001 vs. normoxia. d Western blot analysis of ADORA2A protein expression in HRMECs exposed to hypoxia (0.5% O2) for indicated times. n = 4. HIF-1α and HIF-2α were used as positive controls for hypoxia, and β-actin was used as loading control. e Real-Time PCR analysis of ADORA2A mRNA expression in HRMECs. HRMECs were infected with 10 ( + ) or 30 ( +  + ) pfu per cell of either Ad-mutHIF-1α, Ad-mutHIF-2α, or Ad-Ctrl. n = 3. *P < 0.001. f, g Real-Time PCR analysis of ADORA2A mRNA expression in HRMECs. HRMECs were transfected with siHIF-1α f, siHIF-2α g, or siCtrl. Forty-eight hours later, cells were exposed to hypoxia (0.5% O2) or air (21% O2) for an additional 12 h. n = 4. **P < 0.01. Data are represented as means ± s.e.m. Statistical significance was determined by unpaired Student’s t-test