Anti p50

Western blotting analysis was performed as previously described [48 (link)]. The blot was probed with antibodies specific for iNOS (BD Biosciences), STAT1 (BD Biosciences), pSTAT1 (BD Biosciences), and β-actin (Sigma-Aldrich). Immunoprecipitation was done with anti-p65 and anti-p50 (Santa Cruz Biotech), as previously described [49 (link), 50 (link)]. The immunoprecipitated proteins were analyzed by Western blot analysis with anti-p65 (Santa Cruz Biotech).

Cell culture reagents were obtained from Invitrogen (San Diego, CA, USA), unless otherwise specified. Fetal bovine serum (FBS) was from Biological Industries (Haemek, Israel) and oligofectamine and G418 from Invitrogen. NGF and BDNF from Peprothec (Rocky Hill, NJ). ProNGF from Scil Proteins GmbH (Halle, Germany) and ammonium pyrrolidinedithiocarbamate (PDTC) from Sigma-Aldrich (St. Louis, MO USA). Antibodies used in this study were the following: mouse anti-α-smooth muscle actin (α-SMA, 1∶500; Dako, Dakopatts, Denmark), anti-smooth muscle myosin (1∶400; NeoMarkers, Fremont, CA USA), anti-CD68 (Dako, 1∶250), anti-α-tubulin and anti-Chromosome Region Maintenance 1 (CRM1, 1∶1000; Sigma-Aldrich), rabbit anti-sortilin (1∶200, Abcam, Cambridge, UK; Chemicon Intern, Temecula, CA USA), anti-proNGF (Sigma-Aldrich), anti-bax protein (1∶200), anti-NF-κB p65 (1∶200), anti-p50 (1∶100), anti-IκB-α (1∶50), goat anti-p75^NTR (1∶200), anti-bcl-2 (1∶100, Santa Cruz Biotechnology, CA, USA) and anti-hypoxanthine-guanine phosphoribosyltransferase (HPRT; Abcam). Fluorocrome conjugated secondary antibodies were purchased from Jackson (Suffork, UK) and Invitrogen. Horseradish peroxidase-conjugated secondary antibodies were from Nordic (Tilburg, The Netherlands).

Dulbecco’s modified Eagle’s medium (DMEM), foetal bovine serum (FBS), and other tissue culture reagents were purchased from Gibco BRL Co. (Grand Island, NY). All other chemicals, including lipopolysaccharide (LPS), cobalt protoporphyrin IX chloride (CoPP) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), were obtained from Sigma-Aldrich (St. Louis, MO). Primary antibodies such as anti-iNOS, anti-COX-2, anti-inhibitor kappa B (IкB)-α, anti-p-IкB-α, anti-p50, anti-p65, anti-actin and anti-proliferating cell nuclear antigen (PCNA) antibodies were purchased from Santa Cruz Biotechnology (Dallas, TX). Anti-p-extracellular signal-regulated kinase (ERK), anti-ERK, anti-p-c-Jun N-terminal kinase (JNK), anti-JNK, anti-p-p38, anti-p38, anti-p-protein kinase B (Akt) and anti-Akt were obtained from Cell Signaling Technology (Danvers, MA). Anti-HO-1 antibody was gained from Merck Millipore (Darmstadt, Germany), and anti-Nrf2 antibody was purchased from Abcam (Cambridge, MA). Anti-mouse, anti-goat and anti-rabbit secondary antibodies were supplied by Merck Millipore (Darmstadt, Germany). Tin protoporphyrin IX (SnPP IX), an inhibitor of HO activity, was obtained from Porphyrin Products (Logan, UT). The enzyme-linked immunosorbent assay (ELISA) kit for PGE₂ was purchased from R&D Systems, Inc. (Minneapolis, MN).