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. 2016 Jun 29;30(8):889–904. doi: 10.1210/me.2016-1002

Figure 3.

Figure 3.

Effect of NSIAD V2R mutations on β-arrestin2 recruitment assessed with multiple BRET configurations. HEK293FT cells transiently transfected with cDNA coding for wild-type V2R (black circles), R137C V2R (blue triangles), R137L V2R (red squares), or L312S V2R (orange diamonds) C-terminally tagged with Rluc8 (A) or Nluc (B and C) and β-arrestin2/Venus (A and B) or β-arrestin2/HT (C) were used to determine concentration-response curves with AVP (1pM–10μM) for β-arrestin2 recruitment to wild-type or mutant V2Rs after about 30 minutes. Data are expressed as BRET ratio (above wild-type baseline) as described in Materials and Methods, and the associated kinetic data are shown in Supplemental Figure 1. Data represent mean ± SEM of 5 independent experiments, with curves fitted by nonlinear regression (GraphPad Prism).