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. 2012 Feb 22;32(8):2683–2695. doi: 10.1523/JNEUROSCI.4125-11.2011

Figure 6.

Figure 6.

Neurabin (Nrb) scaffolds formation of the A1R–RGS4 complex. A, The A1R–RGS4 interaction is dependent upon agonist stimulation and the presence of neurabin. Cells coexpressing HA-A1R and RGS4, together with or without Myc-Nrb, were incubated in the presence or absence of R-PIA for 5 min. Cell lysates were subjected to the immunoisolation assay and analyzed using an anti-HA antibody. Representative blots of at least 3 independent experiments are shown. B, RGS4 forms a complex with neurabin and the A1R upon agonist stimulation. Cells coexpressing HA-A1R, RGS4, and Myc-Nrb were incubated with or without R-PIA for 5 min. Cell lysates were subjected to the immunoisolation assay using an anti-RGS4 antibody and analyzed by Western blot. Representative blots of multiple independent experiments are shown. C, R-PIA stimulation fails to promote interaction between neurabin and RGS2 in cells. Cells coexpressing A1R, Myc-Nrb, and RGS2 were stimulated with R-PIA for 5 min. Cell lysates were subjected to immunoisolation assay using an anti-RGS2 antibody and analyzed by Western blot. Representative blots of multiple independent experiments are shown. IP, Immunoprecipitation.