Hplc grade formic acid

Extra pure grade n-hexane and ethyl acetate were purchased from Duksan Pure Chemical (Korea). HPLC-grade formic acid was purchased from Wako (Japan). HPLC-grade acetonitrile was obtained from Fisher Scientific Korea (Korea). Methanol, 3-(4,5-dimethylthiazol-e-yl)-2,5-diphenyl tetrazolium (MTT), propidium iodide (PI) solution, LiCl, crystal violet solution (1%), 37% formaldehyde, and TRI reagent were obtained from Sigma-Aldrich (USA). A RevertAid First Strand cDNA Synthesis Kit and Power SYBR Green PCR Master Mix were purchased from Thermo Fisher Scientific. An Annexin V-FITC/PI Assay Kit and antibodies against cyclin D1 and E2F-1 were purchased from BD Bioscience (USA). Antibodies against WNT2B, c-Myc, Bcl-2, PCNA, and β-actin were purchased from Santa Cruz Biotechnology (USA). Antibodies targeting GSK3α/β, p-GSK3α/β, β-catenin, PARP, CDK4, and HRP-conjugated anti-mouse and anti-rabbit secondary antibodies were obtained from Cell Signaling Technology (USA).

LCMS-grade methanol, HPLC-grade formic acid, and APDSTAG^® Wako Amino Acids internal standard mixture solution composed of 25 stable-isotope labeled amino acids were purchased from FUJIFILM Wako Pure Chemical Corporation (Osaka, Japan). CHCl₃ and LCMS-grade CH₃CN were purchased from Kanto Chemical Co., Inc. (Tokyo, Japan). The other reagents used are mentioned in Supplementary Materials, Section 1.

Dulbecco’s modified Eagle’s minimum essential medium (DMEM), fetal bovine serum (FBS), penicillin, and streptomycin were purchased from Life Technologies Inc. (Grand Island, NY, USA). The antibodies for iNOS, COX-2, p65, PARP, β-actin, normal rabbit IgG and normal mouse IgG_2b were obtained from Santa Cruz Biotechnology Inc. (Santa cruz, CA, USA). The enzyme immunoassay (EIA) kits for PGE₂, IL-6, IL-1β, and TNF-α were purchased from R&D Systems (Minneapolis, MN, USA). TOPscript™ RT Dry MIX was purchased from Enzynomics Co. Ltd. (Daejeon, Korea). iNOS, COX-2, TNF-α, IL-6, IL-1β, and β-actin oligonucleotide primers were purchased from Bioneer (Seoul, Korea). MTT, l-N⁶-(1-iminoethyl)lysine (l-NIL), NS-398, Escherichia coli LPS and all other chemicals were obtained from Sigma Chemical Co. (St. Louis, MO, USA). HPLC-grade acetonitrile was purchased from Fisher Scientific Korea (Seoul, Korea) and HPLC-grade formic acid was purchased from Wako (Osaka, Japan).