Mouse anti inos

Phosphate-buffered saline (PBS) and Dulbecco’s Modified Eagle’s Medium (DMEM) was obtained from Corning, Inc. (New York, NY, USA)., fetal bovine serum (FBS) were obtained from Gibco (Grand Island, CA, USA). Penicillin-Streptomycin was Thermo fisher (Waltham, MA, USA). Lipopolysaccharide (LPS, Escherichia coli 055:B5) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Primary antibodies against rabbit-anti COX-2, NF-κB p65, ERK, phospho-extracellular signal-regulated kinase (p-ERK), p38, p-p38, JNK, phospho-Jun N-terminal (p-JNK), phosphoinositide 3-kinase (PI3K), p-AKT, AKT, and Lamin B1 were purchased from Cell Signaling Technology (Beverly, MA, USA). Mouse anti-iNOS was purchased from from R&D Systems Inc.(Minneapolis, MN, USA). IL-1β and TNF-α were purchased from Cruise Biotechnology (Santa Cruz, CA, USA). Mouse anti-β-actin and goat anti-rabbit IgG HRP antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Water containers were purchased from VWR (Alfragide, Portugal). Raw 264.7 cell line was acquired from American Type Culture Collection (ATCC-TIB-71) and kindly supplied by Dr. Otília Vieira (Center for Neuroscience and Cell Biology, University of Coimbra, Coimbra, Portugal). Dulbecco's Modified Eagle Medium (DMEM) and fetal bovine serum (FBS) were purchased from Thermo Fisher Scientific (Massachusetts, USA). LPS from Escherichia coli (serotype 026:B6) was obtained from Sigma-Aldrich Corp. (St. Louis, MO, USA). The protease and phosphatase inhibitor cocktails were obtained from Roche (Mannheim, Germany). Acrylamide was purchased from Promega (Madison, WI, USA) and the Polyvinylidene difluoride (PVDF) membranes were from Millipore Corp. (Bedford, MA, USA). The primary antibody mouse anti-iNOS was from R&D Systems (Minneapolis, MN, USA) and mouse anti-β-Tubulin was obtained from Sigma-Aldrich Corp. (St. Louis, MO, USA). The anti-mouse Horseradish Peroxidase conjugated secondary antibody was purchased from Santa Cruz Biotechnology (Heidelberg, Germany). Enhanced chemiluminescence (ECL) substrate was obtained from Bio-Rad (Hercules, USA). All other reagents were purchased from Sigma-Aldrich Corp. (St. Louis, MO, USA).