Figure 2. Orthosteric ligand-mediated Nb39 association and dissociation in µ-OR-rHDL.

As described in the Materials and methods, the association and dissociation of Nb39 (1 µM) was measured using an OctetRed^® instrument. Shown is a representative experiment comparing four orthosteric agonists at 30 µM (a) Using GraphPad Prism 6.02, one-phase association lines were fit and the calculated k_obs (b) and k_off (c) for each ligand are plotted (±s.e.m.). Rate constants are means of multiple independent experiments as listed in see Table 1. *Dissociation of Nb39 from the methadone-bound receptor is statistically different from all other ligands (one-way ANOVA with Tukey’s post-hoc test).