5 lox

Antibodies against ERK1/2, phospho-ERK1/2, Akt, phospho-Akt, c-Jun N-terminal kinases (JNK), phospho-JNK, p38, phospho-p38, COX-2, 5-LOX, glyceraldehydes-3-phosphate dehydrogenase (GAPDH), β-actin, lamin b, and Alexa Fluor 594 conjugated goat anti-rabbit IgG antibodies were obtained from Cell Signaling Technology (Boston, MA, USA). Anti-NF-κB and anti-mouse horseradish peroxidase- (HRP-) conjugated IgG antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Dihydroethidium (DHE) was purchased from molecular probes (Eugene, OR, USA). Dexamethasone was obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-rabbit HRP-conjugated IgG secondary antibody and other chemicals were purchased from Sigma-Aldrich (St. Louis, MO, USA), unless otherwise indicated. Gallic acid-l-leucine (GAL) conjugate was synthesized from gallic acid and methyl-l-leucine hydrochloride by following the previous method as described [15 (link)]. The structure of GAL conjugate was confirmed by NMR technique whereas the purity of the product was determined by HPLC analysis, suggesting the purity of up to 98%.

Cultured cells were washed twice in cold phosphate-buffered saline, scraped into lysis buffer and proteins separated on 12% SDS-polyacrylamide gels.^{14 (link)} The following primary antibodies were used: 5-LOX (78 kDa), COX2 (74 kDa), cleaved caspase 1 (20,22 kDa), P-NFkB (70 kDa), NFkB (70 kDa) all from Cell Signaling (Danvers, MA, USA); Beta Amyloid (6E10; Wako, Richmond, VA, USA); actin (45 kDa); (Enzo Life Sciences, Farmingdale, NY, USA); 12-LOX (80 kDa; Cayman, Ann Arbor, MI, USA); 15-LOX (80 kDa; Cayman).

TNF-α, methacholine chloride (MCh), histamine and β-actin antibodies were purchased from Sigma (St. Louis, MO, USA). RvD₂, LTD₄, U-46619, as well as TNF-α and COX-2 antibodies were obtained from Cayman Chemical (Ann Arbor, Michigan). 5-LOX, CysLTR1, P-c-Fos, P-c-Jun, c-Fos, c-Jun, P-p38-MAPK, p38-MAPK antibodies were purchased from Cell Signalling Technology (Boston, MA, USA). DMEM/F-12 and penicillin-streptomycin were obtained from GIBCO Invitrogen Corp. (Burlington, ON, Canada). WRW4 was purchased from Tocris Bioscience (Minneapolis, MN, USA).