Adenosine Receptor A1-A2a Heteromers Regulate EAAT2 Expression and Glutamate Uptake via YY1-Induced Repression of PPARγ Transcription

Figure 2

The A1AR/A2aAR heterodimer regulated the expression of EAAT2 and glutamate uptake. Astrocytes were subjected to OGD conditions in the absence or presence of 30 nM CCPA (an A1AR agonist) and 200 nM SCH58261 (an A2aAR antagonist). (a, b) The expression of EAAT2 and the intracellular glutamate level were detected by western blotting or with a glutamate assay kit, respectively. The data represent as the (). vs. the control group; ^{^} vs. the OGD group; ^# vs. the OGD/CCPA group. Astrocytes were subjected to OGD conditions in the absence or presence of 30 nM CCPA and 200 nM CGS21680 (an A2aAR agonist). (c, d) The expression of EAAT2 and the intracellular glutamate level were detected by western blotting or with a glutamate assay kit, respectively. The data represent as the (). vs. the control group. (e) The protein levels of A2aARafter the transfection of A2aAR siRNA into astrocytes for 48 h were evaluated by western blotting. A2aAR-silenced astrocytes were subjected to OGD conditions in the absence or presence of 30 nM CCPA. (f, g) The expression of EAAT2 and the intracellular glutamate level were detected by western blotting or with a glutamate assay kit, respectively. The data represent as the (). vs. the control group and vs. the OGD group. (h) The protein level of A1AR after transfection of the EYFP-A1AR plasmid into astrocytes for 48 h was evaluated by western blotting. Astrocytes were transfected with the EYFP-A1AR plasmid for 48 h and then subjected to OGD conditions in the absence or presence of 200 nM SCH58261. (i, j) The expression of EAAT2 and the intracellular glutamate level were detected by western blotting or with a glutamate assay kit, respectively. The data represent as the (). vs. the control group; ^{^} vs. the OGD group; ^# vs. the OGD/EYFP-A1AR group.

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