Lipopolysaccharide lps from escherichia coli o111 b4

Fluorescein isothiocyanate (FITC)-conjugated Annexin V/propidium iodide (PI) Kit, anti-CD80, anti-CD86, and phycoerythrin-conjugated anti-MHC-I and anti-MHC-II were purchased from BD Biosciences (San Diego, CA, USA). Lipopolysaccharide (LPS) from Escherichia coli O111:B4 was purchased from InvivoGen (San Diego, CA, USA). Anti-phosphorylated extracellular signal-regulated kinase (ERK)-1/2 monoclonal antibody (Ab), anti-phosphorylated p38 monoclonal Ab, anti-phosphorylated c-Jun N-terminal kinase (JNK) monoclonal Ab, anti-phosphorylated inhibitor of κB (IκB)-α monoclonal Ab, anti-IκB-α monoclonal Ab, anti-nuclear factor (NF)-κB (p65) polyclonal Ab, anti-lamin B polyclonal Ab, anti-Toll-interacting protein (Tollip) monoclonal Ab, anti-suppressor of cytokine signaling (SOCS) 1 polyclonal Ab, anti-interleukin (IL)-1 receptor-associated kinase (IRAK)-M monoclonal Ab, anti-TLR4 polyclonal Ab, anti-cyclooxygenase (COX)-2 polyclonal Ab, anti- inducible nitric oxide (NO) synthase (iNOS) polyclonal Ab, and anti-β-Actin monoclonal Ab were obtained from Cell Signaling Technology (Danvers, MA, USA). Goat anti-rabbit-IgG-horseradish peroxidase (HRP) Ab and goat anti-mouse-IgG-HRP Ab were purchased from Calbiochem (Merck KGaA, Darmstadt, Germany), and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) was purchased from Sigma-Aldrich (St. Louis, MO, USA).

Recombinant mouse granulocyte-macrophage colony stimulating factor (rmGM-CSF) and recombinant mouse interleukin (rmIL)-4 were purchased from JW Creagene (Korea). Roswell Park Memorial Institute (RPMI) 1640 medium, fetal bovine serum (FBS), and penicillin/streptomycin were obtained from Gibco (Grand Island, USA). Lipopolysaccharide (LPS) from Escherichia coli O111:B4 (ultra-pure grade) was obtained from InvivoGen (USA). V450-conjugated anti-CD11c, fluorescein isothiocyanate (FITC)-conjugated anti-CD80, anti-IL-10, anti-CD4, annexin V/propidium iodine (PI) kit, phycoerythrin (PE)-conjugated anti-CD86, anti-IL-12p70, allophycocyanin (APC)-conjugated tumor necrosis factor (TNF)-α, PE-Cy7-conjugated anti-CD11c, anti-CD8, BV510-conjugated live/dead kit, and enzyme linked immunosorbent assay (ELISA) kits for TNF-α, IL-12p70, IL-10, IFN-γ, IL-2, and IL-5 were purchased from BD biosciences (USA). APC-conjugated anti-MHC-I, PE-Cy7-conjugated anti-MHC-II, anti-CD8, 2-mercaptoethanol, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) buffer, Minimum Essential Medium (MEM) non-essential amino acids, and CellTrace Violet Cell Proliferation Kit were purchased from Invitrogen (USA).

Recombinant mouse granulocyte-macrophage colony stimulating factor (GM-CSF) and interleukin (IL)-4 were purchased from JW CreaGene (Daegu, Korea). Lipopolysaccharide (LPS, from Escherichia coli O111:B4) were purchased from Invivogen (San Diego, CA, USA). The annexin V/propidium iodide (PI) kit was purchased from BD Bioscience (San Diego, CA, USA). ELISA kits for TNF-α, IL-6, IL-1β, IL-4, IL-12p70, and IL-10 were purchased from eBioscience (San Diego, CA, USA). The OT-I (OVA_257−264) and OT-II (OVA_323−339) peptides were synthesized by Abfrontier (Seoul, Korea). Fluorescein-conjugated antibodies (Abs) to, MHC-I, MHC-II, T-bet, and GATA-3 were purchased from eBioscience. Fluorescein-conjugated Abs to, CD11c, CD80, CD86, TNF-α, IL-12p70, IL-10, CD4, IFN-γ, IL-4, and CD8α were purchased from BD Bioscience. Anti-phosphorylated ERK1/2, -phosphorylated JNK, -phosphorylated p38, -phosphorylated IκB-a, IκB-a, NF-κB (p65), and lamin B were purchased from Santa Cruz Biotechnology (CA, USA). Anti-β-actin mAb (AC-15) and FITC-dextran (molecular mass, 40 kDa) were purchased from Sigma (St. Louis, MO, USA). All the pharmacological inhibitors were purchased from Calbiochem (San Diego, CA, USA).