*5.10. Quantification of Cytokines and Chemokines Produced by THP-1 Macrophages*

The cytokines and chemokines produced by THP-1 macrophages in response to the treatments with BaV and Kn-Ba were evaluated in cell-free supernatants. For quantification of inflammatory cytokines IL-1β, IL-6, IL-10, IL-12, and TNF-α, the CBA Human Inflammatory Cytokines kit was used (BD, Bioscience, Franklin Lakes, NJ, USA), and for quantification of the chemokines CXCL8/IL-8, CCL5/RANTES, CXCL9/MIG, CCL2/MCP-1, and CXCL10/IP-10, the CBA Human Chemokines kit was used (BD, Bioscience, Franklin Lakes, NJ, USA), according to the manufacturer's instructions. The data acquisition was performed in a flow cytometer (FACS Aria III, Becton Dickinson, San Jose, CA, USA) and analyzed by the software FACS Diva, version 6.1.3.

#### *5.11. Quantification of Lipid Inflammatory Mediators Produced by THP-1 Macrophages*

The concentration of the lipid mediators LTB4 and PGE2 in cell-free supernatants was quantified by competitive ELISA using specific kits (EIA, Cayman Chemical, Ann Arbor, MI, USA), according to the manufacturer's instructions. The absorbances were determined in a spectrophotometer at λ 405/420 nm (VersaMax, Molecular Devices, San Jose, CA, USA). The eicosanoids concentrations were calculated by the interpolation on the kits' standard curve.
