Figure 8.

Characterization of [³H]140 binding. Association (a) and dissociation (b) kinetics of specific binding of 1.02 nM [³H]140 to membranes of Flp-In TREx 293 cells induced to express a human GPR84-Gα_i2 fusion protein are shown from representative experiments. Dissociation was assessed in the presence of 1 μM compound 020 to prevent reassociation of the radioligand. Subsequently, the specific binding of a range of concentrations of [³H]140 to membranes of Flp-In TREx 293 cells induced to express the human GPR84-Gα_i2 fusion protein was measured (c). Similar studies were performed on membranes expressing Ala¹⁰²Thr-Gly³⁶³Ser human GPR84-Gα_i2 (d), mouse GPR84-Gα_i2 (e), and Thr¹⁰²Ala-Ser³⁶³Gly mouse GPR84-Gα_i2 (f). Specific binding with estimated K_d 4.7 ± 0.8 nM was observed to Thr¹⁰²Ala-Ser³⁶³Gly mouse GPR84-Gα_i2.