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. Author manuscript; available in PMC: 2013 Jan 1.
Published in final edited form as: J Neurochem. 2011 Nov 24;120(1):56–69. doi: 10.1111/j.1471-4159.2011.07559.x

Fig. 3. Effect of D2R co-expression on the TX100 solubility of RGS9-2 and Gβ5 co-expressed in HEK293T cells and effect of dopamine (DA)-treatment on D2R-mediated translocation of RGS9-2 to TX100-insoluble cellular fractions.

Fig. 3

A. HEK293T cells were transfected with cDNA for RGS9-2-V5-tagged (RGS9) and Gβ5 or with cDNA for RGS9, Gβ5 and D2R. Western blot (WB) of RGS9-2-V5 (RGS9) in TX100 detergent soluble (S) and insoluble (P, pellet) cell fractions probed with an anti-V5 epitope-tagged antibody, as described in the Materials and Methods. B. Levels of RGS9-2 protein in TX100-soluble (closed black bars) and insoluble (open bars) cellular fractions visualized in A and expressed as percentage of the total expressed RGS9-2 protein (mean ± S.D., n=4, *represents p<0.01 compared to RGS9-2 cDNA/Gβ5 cDNA transfected group). C. Western blot (WB) of Gβ5 in TX100 detergent soluble (S) and insoluble (P, pellet) cell fractions described in A. D. Levels of Gβ5 protein in TX100-soluble (closed black bars) and insoluble (open bars) cellular fractions visualized in C and expressed as percentage of the total expressed Gβ5 protein (mean ± SEM, n=4, *represents p<0.01 compared to RGS9-2 cDNA/Gβ5 cDNA transfected group). E. HEK293T cells transfected with expression plasmids for D2R, Gβ5 and a RGS9-2 were treated for 10 or 30 min with 1 μM dopamine (DA). RGS9-2 proteins in TX100 detergent soluble (S) and insoluble (P, pellet) fractions from cells treated with DA as indicated and visualized by immunoblotting. F. Levels of RGS9-2 protein in TX100-soluble (closed black bars) and insoluble (open bars) cellular fractions visualized in E and expressed as percentage of the total cellular RGS9-2 protein (mean ± SEM, n=4).