S 5 adenosyl l methionine p toluenesulfonate salt

For DNA transfection in HEK293T cells, transfection grade linear polyethylenimine (PEI) hydrochloride was purchased from Polysciences, Inc. S-(5′-Adenosyl)-L-methionine p-toluenesulfonate salt and cytidine 5′-triphosphate disodium salt hydrate were purchased from Sigma Aldrich. POPC (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine,) and POPE (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine) were purchased from Avanti Polar Lipids. Amberlite XAD-2 hydrophobic beads, Anti-FLAG M2 magnetic beads, and 3X FLAG peptide were purchased from Millipore Sigma. Pierce silver stain kit was purchased from ThermoFisher Scientific. Rabbit polyclonal viperin antibody (11833–1-AP) was purchased from ProteinTech. Goat anti-rabbit Ig secondary antibody (170-6515) was purchased from BioRad.

Methanol, acetonitrile, ammonium formate, glucose-6-phosphate, β-nicotinamide adenine dinucleotide phosphate hydrate (NADP⁺), uridine 5′-diphosphoglucuronic acid trisodium salt (UDPGA), adenosine 3′-phosphate 5′-phosphosulfate triethylammonium salt (PAPS), S-(5′-adenosyl)-L-methionine p-toluenesulfonate salt (SAM), magnesium chloride hexahydrate and sodium citrate tribasic dihydrate were purchased from Sigma-Aldrich (St. Louis, MI, USA). Formic acid was acquired from Scharlab (Sentmenat, Barcelona, Spain) and Gentest™ 0.5 M phosphate buffer pH 7.4 and glucose-6-phosphate dehydrogenase from Corning (Woburn, MA, USA). Ultrapure water was obtained using a Mili-Q RG system from the company Milipore (Burlington, MA, USA). NEP hydrochloride reference material (98% purity) was purchased from Cayman Chemical (Ann Arbor, MI, USA) and a stock solution prepared at 1 mg/mL (4.1 mM free base) in Methanol. NEP working solutions of 40 and 732 μM were also prepared by appropriate dilution of NEP stock solution in Methanol. Dapaconazole, the positive control for the experiment, was supplied by Biolab Farmacêutica Ltda. (São Paulo, Brazil) and a stock solution prepared at 1 mg/mL (2.4 mM) in Methanol. A dapaconazole working solution of 40 μM was prepared from a dapaconazol stock solution in Methanol.