Bay11 7028

LPS derived from Escherichia coli strain 055:B5, Trizol, Lipofectamine 2000 and BAY11-7028, the inhibitor of nuclear factor kappa B (NF-κB) and aspirin were purchased from Sigma (St. Louis, MO, USA).

LPS derived from Escherichia coli strain 055: B5, and the NF-κB inhibitor BAY 11–7028 were purchased from Sigma-Aldrich. Recombinant human TNF-α was obtained from PeproTech. Poly (I:C) was obtained from InvivoGen. The following antibodies were used for western blotting: an anti-TLR4 rabbit polyclonal antibody (Abcam); anti-TLR3 rabbit polyclonal antibody (Cell Signaling Technology); anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) rabbit polyclonal antibody (Bioworld Technology); anti-HIF-1α rabbit polyclonal antibody (Cell Signaling Technology), anti-VEGF rabbit polyclonal antibody (Abcam). Protein standards were obtained from SunShineBio. Horseradish peroxidase (HRP)-conjugated goat anti-rabbit immunoglobulin G (IgG) was purchased from Immunology Consultants Laboratory (Newberg, OR, USA). An anti-p65 rabbit polyclonal antibody (Cell Signaling Technology) and goat anti-rabbit IgG (Bioworld Technology) were used for immunofluorescence.

RPMI 1640 medium, DMEM (Dulbecco’s Modified Eagle Medium), fetal bovine serum (FBS), and a 0.02% ethylenediaminetetraacetic acid (EDTA) solution were obtained from Invitrogen (Gibco BRL, Grand Island, NY, USA). The human B7-H3 antibody, recombinant human B7-H3 protein, human B7-H3 ELISA kit, and the IL-8 and VEGF immunoassay ELISA kits were purchased from R&D Systems (Minneapolis, MN, USA). The PGL4.32 (Luc2p/NF-κB-RE/Hygro) luciferase reporter vector, the PRL-TK vector dual-luciferase reporter assay system and the FuGENE 6 Transfection Reagent were purchased from Promega (Madison, WI, USA). The anti-TLR4 and anti-NF-κB p65 antibodies were obtained from Abcam (Cambridge, UK), the NF-kB inhibitor BAY11-7028 was obtained from Sigma (St. Louis, MO, USA), and the TLR4 inhibitor TAK-242 was purchased from InvivoGen (San Diego, CA, USA).