Fig. 1.

Differential agonist-dependent receptor activation kinetics and dynasore sensitivity of HCA₃ and GPR84 DMR responses. CHO-K1 cells were transiently transfected with receptor constructs. a cAMP inhibition assays in presence of 2 μM forskolin (fsk) were performed which showed activation of HCA₃ by 3-hydroxyoctanoic acid (3HO) and 3-hydroxydecanoic acid (3HDec). C10 and 3HDec activated GPR84. cAMP level of HCA₃- or GPR84-transfected cells in absence of agonist is set 100%, respectively. b Transfected CHO-K1 cells were seeded in fibronectin-coated Epic plates and DMR responses were recorded. 3HO and 3HDec induced a pertussis toxin (PTX, 100 ng/ml)-sensitive DMR response in HCA₃-, C10 and 3HDec in GPR84-expressing CHO-K1 cells. DMR responses in absence of dynasore showed distinct receptor activation kinetics for both, HCA₃ and GPR84, when comparing the shown agonists, respectively. Dynasore (80 µM) diminished the 3HDec-induced DMR response of HCA₃-expressing cells completely. Time points 10 min, 20 min and 40 min were used to generate bar graphs (concentration-response curves see Figures S2, S3) to highlight, that DMR responses of HCA₃ to 3HO and GPR84 to 3HDec were affected by dynasore only at earlier time points. The DMR responses of HCA₃ to 3HDec and of GPR84 to C10 were diminished over the whole time recorded. Shown is the agonist-induced wavelength shift in pm of three to five independent experiments, each carried out in triplicates as mean ± SEM. ns, not significant; ^#P ≤ 0.1; * P ≤ 0.05; ** P ≤ 0.01