Addendum: Resolving Inflammation: Synthesis, Configurational Assignment, and Biological Evaluations of RvD1n−3 DPA.

(1) Ligand receptor interactions were monitored using the PathHunter SP ® sp -arrestin cell-based assays (Eurofins DiscoverX Corporation, Fremont, CA, USA) essentially as described earlier. SP [1] sp CHO- -arrestin-GPR32 cells and HEK- -arrestin-FPR2/ALX cells were purchased from DiscoverX, and maintained in HAM F-12 and DMEM media, respectively, supplemented with 10% FBS, 1X pen/strip, G418 (0.8 mg ml SP -1 sp ) and hygromycin (0.3 mg ml SP -1 sp ). (5) DiscoverX PathHunter Detection Reagent (Eurofins DiscoverX Corporation, Fremont, CA, USA) was then used following the manufacturer's instruction; namely, 55 microliters of Working Detection Solution (mixture of Cell Assay Buffer: Substrate Reagent 1: Substrate Reagent 2; 19:5:1) was added to each well, and assay plates incubated in the dark at room temperature for 60 min. Note that the receptor denoted ALX/FPR2 was demonstrated bias in intracellular signaling with specific ligands and with pro-resolving mediator ligands, SP [2] sp confirmed in >175 publications in PubMed to date. [Extracted from the article]