Human macrophages in 12-well plates were incubated for 7 hours at 37°C with vehicle or 10 nM γ-carboxylated Gas6. The cell culture medium was then harvested for analysis by (i) the Neogen LXA4 ELISA kit, which has the following cross-reactivities according to the manufacturer: 15-epi-LXA4 (24%), 5(S),6(R)-DiHETE (5%), LXB4 (1%), and 15-HETE (0.1%); (ii) the Cayman LTB4 ELISA kit, which has cross-reactivities to 5,6-DiHETE (0.07%), 5(R)-HETE (3.7%), 15(R)-HETE (0.98%), 15(S)-HETE (0.4%), 5(S)-HETE (6.6%), 20-hydroxy LTB4 (2.7%), 6-trans-12-epi LTB4 (0.31%), and 6-trans LTB4 (0.11%); or (iii) the Cayman RvD1 ELISA kit, which has cross-reactivities to 5(S),6(R)-LXA4 (20%), 17(R)-RvD1 (4.2%), and 10(S),17(S)-DiHDoHE (0.7%). We refer to these lipid mediators as iLXA4, iLTB4, and iRvD1, respectively, where “i” stands for “immunoreactive.” The data are reported as fold change in abundance in the experimental group relative to that in the control group, which was set at 1.0. Similar analyses were conducted on peritoneal exudates for the zymosan experiments. In all cases, 50 μl of sample was assayed as per the manufacturer’s instructions.

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