Cxcl9 mig

The chemokines were dissolved in water for injection. CXCL1/KC and CXCL5/LIX were obtained from Sigma Aldrich and CXCL9/MIG and CXCL12/SDF-1 were from R&D System. Reconstituted chemokines were injected in the following concentrations: 10 ng/5 μL, 100 ng/5 μL, and 500 ng/5 μL. The behavioral tests were performed after 1, 4, and 24 hours following chemokine administration.

Total blood samples were collected into vacutainer tubes (cat. no. 366881, BD Biosciences, Plymouth, UK), allowed to clot for 1 h at 37 °C, and centrifuged for 15 min at 1700 rcf at 4 °C. Then, serum was aliquoted and stored at − 80 °C until use. Concentrations of human CD25/IL-2 Rα (cat. no. DY223, R&D Systems, Minneapolis, USA), CD27/TNFRSF7 (cat. no. DY382-05, R&D Systems, Minneapolis, USA), IL-17A (cat. no. DY317, R&D Systems, Minneapolis, USA), S100B (cat. no. DY1820-05, R&D Systems, Minneapolis, USA), CXCL9/MIG (cat. no. DY392, R&D Systems, Minneapolis, USA) and CXCL11/I-TAC (cat. no. DY672, R&D Systems, Minneapolis, USA) were measured using DuoSet ELISA kits (R&D Systems, Minneapolis, USA). CXCL10/IP-10 levels were evaluated with a BD Pharmingen kit (cat. no. 550926, San Diego, USA). The plates were analyzed in an ELISA iMark Microplate Reader (Bio-Rad, California, USA).