Fig. 2. Mutations of T³⁴⁷ and S³⁵⁰ do not significantly affect βarr recruitment and trafficking.

(A to D) Mutations of either T³⁴⁷ or S³⁵⁰ do not significantly affect agonist-induced (100 nM AVP) βarr recruitment as assessed by co-IP experiment in HEK-293 cells. Representative images from four independent experiments (five for S³⁵⁰ + βarr2), and densitometry-based quantification of data, normalized with respect to βarr co-IP for the V₂R^WT at 30-min agonist stimulation (treated as 100%), are shown. Data are analyzed using two-way ANOVA. (E and F) Agonist-induced trafficking of βarrs for the V₂R^T347A and V₂R^S30A is similar to that of V₂R^WT as assessed by confocal microscopy. HEK-293 cells expressing the indicated receptor mutant and βarr-mYFP were stimulated with 100 nM AVP, and representative images from three independent experiments at indicated time points are shown. Scale bars, 10 μm.