Figure 9.

β-arrestin 2^{K18R, K107R, K108R, K207R, K296R}demonstrates impaired agonist-promoted co-localiztion with M₂ mAChR in MEF KO1/2. MEF KO1/2 cells were transfected with HA-M₂ mAChRs (green) and either wild-type FLAG-β-arrestin 2, FLAG-β-arrestin 2^{K18R, K107R, K108R, K207R, K296R} or FLAG-β-arrestin 2^{K11R, K12R} (red). 24 hr after transfection, cells were treated with 1 mM carbachol for 30 min. Cells were fixed, probed, and imaged as described in methods. Wild-type and β-arrestin 2^{K11R, K12R} demonstrate co-localization with the M₂ mAChR which is significantly diminished with β-arrestin 2^{K18R, K107R, K108R, K207R, K296R}. Image shown is representative of two independent experiments.