Specific primers for real time pcr

HT-29 (HTB-38) and THP-1 (TIB-202) cells were purchased from the American Type Culture Collection (ATCC). RPMI-1640, D-MEM, fetal bovine serum (FBS), L-glutamine, HEPES, sodium pyruvate, glucose, 2-mercaptoethanol, penicillin/streptomycin mixture, phosphate buffered saline solution (PBS), mouse monoclonal antibody against β-actin, horseradish peroxidase (HRP)-conjugated anti-mouse and anti-rabbit secondary antibodies and other chemicals of analytical grade were from Sigma-Aldrich (Milan, Italy). Lipopolysaccharide (LPS) was from InvivoGen (San Diego, CA, USA). Rabbit monoclonal antibody for nuclear factor erythroid 2-related factor 2 (Nrf2) was from Bioss Antibodies (Aurogene Srl, Rome, Italy). The rabbit antibodies against NOS2 and laminin B1 were purchased from Abcam (Cambridge, UK). Mouse MMP-2 polyclonal antibody was obtained from Chemicon International Inc. (Merck, Milan, Italy). Mouse monoclonal antibodies against SOD2 were from Santa Cruz Biotechnology (D.B.A. Italia S.R.L., Milan, Italy). Pierce ECL Western Blotting Substrate, TRIzol, the high-capacity cDNA archive kit and SYBR Select Master mix were from Life Technologies (Milan, Italy). Specific primers for real-time PCR were from Eurofins Genomics (Ebersberg, Germany). ELISA kits for the quantitative detection of human IL-1β and TNF-α were from Cloud-Clone Corp. (Katy, TX, USA).

The human colonic epithelial cell line, HT-29, was purchased from Sigma Aldrich (Milan, Italy). RPMI-1640, L-glutamine, HEPES, sodium pyruvate, glucose, 2-mercaptoethanol, penicillin/streptomycin mixture, dimethyl sulfoxide (DMSO), lipopolysaccharide (LPS), 3-(4,5-methylthiazol-2-yl)-2,5-diphenyl-tetrazoliumbromide (MTT), phosphate buffered saline solution (PBS), was from InvivoGen (San Diego, CA, USA). Fetal bovine serum (FBS), as well as TRIzol, High-capacity cDNA archive kit, SYBR Select Master Mix, the biotin 3′-end DNA labeling kit, LightShift Chemiluminescent Electrophoretic Mobility Shift assay (EMSA) kit, Biodyne Nylon membranes, and Super-Signal West Pico chemiluminescent substrate system were from Life Technologies (Milan, Italy). Specific primers for Real-Time PCR were from Eurofins Genomics (Ebersberg, Germany). ELISA kits for the quantitative detection of human TNF-α and IL-1β were from Cloud-Clone Corp. (Katy, TX, USA). BA was provided by Enfarma SRL (Misterbianco, Italy) [13 (link)].
BA (SB root dry extract titrated at 95% in BA) was dissolved in DMSO in a 10 mg/mL stock solution, stored at −20 °C, and diluted to different concentrations with culture medium right before experimental use.