Figure 2 – Selection of the best G_s-biased candidates.

Multivariate analysis of (A) maximal efficacies (E_max) extrapolated from curve fitting of the concentration/activity curves from Fig. 1 and Table S1 and (B) transduction coefficients (TC) obtained after statistical fitting of the operational model of agonism to data extrapolated from Fig. 1, were represented in radial plots. E_max range is 0 – 100 while TC range is 0 – 10. Note that values that were very low and could not be accurately determined were given a value of zero in these plots. Direct comparison of concentration/activity curves for Gα_s recruitment and β-arrestin recruitment induced by ISO, RAC, DOB and HIG at the β₂AR (C) and the β₁AR (D). Data for β₂AR comparison were taken from Fig. 1A and B. For β₁AR comparison, HEK293 cells were transiently transected with BRET donor β₁AR-Rluc and BRET acceptor NES-venus-mGs or GFP-β-arrestin 2 depending on the read-out. After 48 h, cells were stimulated with increased concentration of ISO, RAC, DOB or HIG (10⁻¹⁰ to 10⁻⁴ M) for 20 min. Signal generated by each condition was plotted and fitted using the three parameters non-linear regression curve fitting in GraphPad Prism to generate concentration/activity curves. Signal was normalized to the average maximal response of ISO as 100%. Data are mean ± SEM, n = 6.