Poly adp ribose polymerase parp 9542

Inducible nitric oxide synthase (iNOS; M00368) was used as a primary antibody from Boster Biological Technology (Pleasanton, CA, USA). Inhibitors of κB-α (IκB-α; sc-203), IκB kinase-α/β (IKK-α/β; sc-7607), HO-1 (sc-136960), and β-actin (sc-47778) were used as primary antibodies (Santa Cruz Biotechnology, Inc., Dallas, Texas, USA). NF-κB p65 (4764), phospho-IκB-α (9246), phospho-IKK-α/β (Ser176/180; 2697), kelch-like ECH-associated protein 1 (keap1; sc-514914), and poly ADP-ribose polymerase (PARP; 9542) were used as primary antibodies (Cell Signaling Technology, Danvers, MA, USA). Nrf2 (ab92946) was used as a primary antibody (Abcam, Cambridge, UK). Horseradish peroxidase-conjugated (HRP) secondary antibodies were obtained from Jackson ImmunoResearch (West Grove, PA, USA). Commercial enzyme-linked immunosorbent assay (ELISA) kits for PGE2, myeloperoxidase (MPO), and SOD were purchased from R&D Systems (Minneapolis, MN, USA), Biovision (Milpitas, CA, USA), and Abcam (Cambridge, MA, USA). Unless otherwise specified, all remaining reagents were purchased from Sigma-Aldrich (St. Louis, MO, USA).

Stationary phases and eluents used in column chromatography and extraction were: silica gel (mesh 0.063–0.2 mm, Merck, Darmstadt, Germany); polyamide (Roth, Karlsrue, Germany); n-hexane, diethyl ether, benzene, and chloroform (Chempur, Pekary Śląskie, Poland); methanol, ethyl acetate, and acetone (POCH, Gliwice, Poland).
Antibody against β-actin (A2066, dilution 1:2000), horseradish peroxidase-conjugated secondary antibodies, anti-rabbit (A9169, dilution 1:5000) and anti-mouse (A9044, dilution 1:5000), and 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide (MTT) were obtained from Sigma-Aldrich (Poznań, Poland). Dulbecco’s modified Eagle’s medium (DMEM), penicillin, streptomycin, fetal bovine serum (FBS), phosphate-buffered saline, trypsin, and Hoechst 33342 were purchased from Thermo Fisher Scientific (Warszawa, Poland). Antibodies against caspase-3 (9662, dilution 1:1000), cleaved caspase-3 (9664, dilution 1:1000), caspase-7 (12827, dilution 1:1000), caspase-8 (9746, dilution 1:1000), caspase-9 (9508, dilution 1:1000), Bcl-2 protein family member, BID (2002, dilution 1:1000), poly(ADP-ribose) polymerase (PARP; 9542, dilution 1:1000) and cleaved PARP (5625, dilution 1:1000) were obtained from Cell Signaling Technology (Danvers, MA, USA). Antibodies against p53 (sc-126, dilution 1:1000) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).