Fig. 3
Blockade of A2AR decreases the expression and immunoreactivity of iNOS and NO production induced by LPS or EHP. Retinal organotypic cultures were pretreated with SCH 58261 (50 nM) and then challenged with LPS (3 μg/mL) or EHP (+70 mmHg) for 4 h. a iNOS mRNA expression was assessed by qPCR. Results are presented as fold change of the control, from six to twelve independent experiments. b Organotypic retinal cultures were immunostained for iNOS (green) and CD11b (microglia marker; red) and imaged in the GCL using a confocal microscope. Nuclei were stained with DAPI (blue). The images are representative of four to five independent experiments. c The immunoreactivity of iNOS in microglia localized in the GCL was quantified. Results are expressed as percentage of control from four to five independent experiments. d The production of NO was assessed by the Griess reaction in the culture supernatants, and nitrite formation was quantified. Results are expressed as percentage of control and are mean ± SEM of four to six independent experiments. *P < 0.05, **P < 0.01 and ***P < 0.001, different from control; + P < 0.05, different from LPS; # P < 0.05 and ### P < 0.001, different from EHP; Kruskal-Wallis test, followed by Dunn’s multiple comparison test