FLA-ST Ultrapure (purified flagellin from Salmonella Typhimurium, a TLR5 agonist) and the vehicle solution, endotoxin-free water, were purchased from InvivoGen (San Diego, CA, USA). In this experiment, it was applied at 0.1 μg, 0.3 μg and 0.9 μg in 10 μL vehicle solution separately, which was modified from the previous procedure of intraplantar administration with flagellin [19 (link)].
Go 6976 was purchased from Tocris Bioscience (Ellisville, MO, USA) and prepared in 0.1% DMSO in 0.1 M PB [57 (link)]. FLA-ST Ultrapure 0.9 μg (InvivoGen, San Diego, CA, USA) was used to combine with Go 6976, which separated into four subgroups (0 μM, 5 μM, 20 μM and 50 μM; n = 3 per subgroup) respectively. Likewise, DAMGO 25 μg (Tocris Bioscience, Ellisville, MO, USA) was used to combine with Go 6976 and also assigned into four subgroups separately.
Go 6976 was purchased from Tocris Bioscience (Ellisville, MO, USA) and prepared in 0.1% DMSO in 0.1 M PB [57 (link)]. FLA-ST Ultrapure 0.9 μg (InvivoGen, San Diego, CA, USA) was used to combine with Go 6976, which separated into four subgroups (0 μM, 5 μM, 20 μM and 50 μM; n = 3 per subgroup) respectively. Likewise, DAMGO 25 μg (Tocris Bioscience, Ellisville, MO, USA) was used to combine with Go 6976 and also assigned into four subgroups separately.
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