Pd184352

5Z-7-oxozeaenol, NG25, BI605906, BIRB0796, and C1 were kindly provided by Professor Sir Philip Cohen (MRC PPU, University of Dundee, UK). PD184352 was bought from US Biological (Swampscott, MA, USA). Flagellin, Pam3CSK4, LPS, and the polyclonal antibodies against TLR2 (cat #pab-hstlr2), TLR4 (cat #pab-hstlr4), and TLR5 (cat #pab-hstlr5) were purchased from InvivoGen (San Diego, CA, USA). Antibodies against p44/42 ERK MAPK (cat #9107) and phospho-ERK Thr²⁰²/Tyr²⁰⁴ (cat #4370) were purchased from Cell Signaling Technology. Recombinant human IL-1β was purchased from R&D systems (Minneapolis, MN, USA). The clinical isolate of Pseudomonas aeruginosa is the same isolate used previously (Bérubé et al., 2010 (link)). CnT-BM.1 cell culture media and A, B, and C supplements (CnT-17.S) were purchased from CELLnTEC (Bern, Switzerland). Nuclear/cytoplasmic extract kit was purchased from Active Motif (Carlsbad, CA, USA, cat #40010).

To determine the pathways involved in IL-17 induced B cell migration, we used pharmacologic inhibitors directed against various pathways that are potentially known to be activated by IL-17 and may be involved in cells migration. B cells (50×10³) were incubated with the inhibitors for 1 hour at 37°C prior to migration assay. Specifically, we used the p38 MAPK inhibitor SB203580 (0.1 mM; Axon Medchem, Groningen, The Netherlands), the extracellular signal-regulated kinase (ERK) 1/2 MAPK inhibitor PD184352 (2 mM; United States Biological, Inc, Swampscott, Mass), the NF-kB inhibitor PS1145 (10 mM; Professor Sir Philip Cohen), and the phosphoinositide 3-kinase (PI3K) inhibitor PI103 (5 mM; Cayman Chemical, Ann Arbor, Mich). All results were compared with the corresponding vehicle control dimethyl sulfoxide.