Fig. 5 | Nature Communications

Fig. 5

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

Fig. 5

Adenosine-ADORA2A signaling regulates glycolysis in HRMECs and mouse retinal ECs. a Scheme showing the glycolytic pathway and associated enzymes. b Real-Time PCR analysis of the mRNA levels of glycolytic genes in HRMECs transfected with siA2AR or siCtrl under normoxia (21% O2) or hypoxia (0.5% O2). n = 5. *P < 0.05, **P < 0.01, ***P < 0.001 vs. siCtrl normoxia group; & P < 0.05, && P < 0.01, &&& P < 0.001 vs. siCtrl hypoxia group. c Real-Time PCR analysis of the mRNA levels of glycolytic genes in retinal blood vessels isolated with laser-capture microdissection from OIR-Adora2a WT and Adora2a VEC-KO mice at P17. n = 4. *P < 0.05 vs. Adora2a WT group. d Levels of secreted lactate of HRMECs transfected with siA2AR or siCtrl under normoxia or hypoxia for indicated times. n = 3. ***P < 0.001. e ECAR profile showing glycolytic function in siCtrl- and siA2AR-transfected cells under hypoxia (0.5% O2) or normoxia (21% O2). Vertical lines indicate the time of addition of glucose (10 mmol/l), oligomycin (2 μmol/l), and 2-DG (50 mmol/l). f Quantification of glycolytic function parameters from e. n = 8 for normoxic groups and n = 16 for each of CoCl2 treatment groups. *P < 0.05; ***P < 0.001. g Real-Time PCR analysis of the mRNA levels of glycolytic genes in HRMECs infected with Ad-Ctrl or Ad-A2AR, with or without adenosine treatment. n = 4. *P < 0.05; **P < 0.01; ***P < 0.001 vs. Ad-Ctrl; & P < 0.05; && P < 0.01; &&& P < 0.001 vs. Ad-Ctrl + Ado. h Levels of secreted lactate of HRMECs infected with Ad-A2AR or Ad-Ctrl with or without adenosine treatment. n = 3. ***P < 0.001 vs. Ad-Ctrl. i ECAR profile showing glycolytic function in Ad-Ctrl- and Ad-A2AR-infected cells, with or without adenosine treatment. j Quantification of glycolytic function parameters from i. n = 8 per group. *P < 0.05; **P < 0.01; ***P < 0.001 vs. Ad-Ctrl. Data are represented as means ± s.e.m. Statistical significance was determined by unpaired Student’s t-test (for b, c, g), one-way ANOVA followed by Bonferroni test (for f, j), and two-way ANOVA followed by Bonferroni test (for d, h)