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. 2018 Mar 20;12:187. doi: 10.3389/fnins.2018.00187

Figure 2.

Figure 2

D2R forms complexes with A2AR in NSC34. (A) NSC34 cells were transfected with FLAG-A2AR and V5-D2R for 48 h. Next, cells were stained with a FLAG antibody (red), a V5 antibody (green) and nuclear marker (DAPI, blue). Scale bar: 10 μm. (B) To verify the interaction between FLAG-A2AR and V5-D2R, cells were stained with a FLAG antibody and a V5 antibody by using the PLA detection method. The cell morphology was analyzed using Rhodamine-phalloidin staining (red). Scale bar: 10 μm. (C) NSC34 cells were transfected with the indicated plasmids for 48 h. Next, cells were lysed to examine the interaction between FLAG-A2AR and V5-D2R by immunoprecipitation (IP) using the indicated antibodies. (D) NSC34 cells were incubated with T1-11 (30 μM) in the absence or presence of quinpirole (QP; a D2R agonist, 1 μM) for 15 min. Cells were harvested to determine cAMP production. (E) NSC34 cells were treated with T1-11 (30 μM) in the absence or presence of QP (1 μM) for 30 min. Next, cells were harvested to determine PKA activity. *p < 0.05, significantly different between the indicated groups. Data are presented as the mean ± SEM of three independent experiments.