Heneicosanoic acid

Dulbecco’s Modified Eagle Medium/Ham’a F12 (DMEM/F12 1:1) and bovine calf serum were purchased from Life Technologies Corporation (Carlsbad, CA, USA). Ketoconazole, Indomethacin, SQ 22536, H-89, lipopolysaccharides from Escherichia coli 055:B5 and forskolin were purchased from Sigma-Aldrich (St. Louis, MO, USA), and normal goat serum was acquired from Dako (Carpentaria, CA, USA). Anti-Iba-1 antibody and anti-glial fibrillary acidic protein (GFAP) antibody were purchased from Wako (Richmond, VA, USA) and Sigma-Aldrich (St. Louise, MO, USA), respectively. Antibodies to PKA, CREB, and GAPDH were purchased from Cell Signaling. Heneicosanoic acid was from Nu-Chek Prep (Elysian, MN). Butylated hydroxytoluene (BHT), LY311727, neomycin sulfate, and 10% boron trifluoride-methanol (BF₃/methanol) were from Sigma-Aldrich (St. Louis, MO). Methyl arachidonyl fluorophosphonate (MAFP), bromoenol lactone (BEL), and ZLJ-6 were obtained from Cayman Chemical (Ann Arbor, MI). 2-TEDC was from Tocris Bioscience (Bristol, UK). HPLC grade water and acetonitrile were from Avantor Performance Materials (Center Valley, PA). Other solvents, BCA protein assay kit, and acetic acid were from Fisher Scientific (Pittsburgh, PA)

High-performance LC grade solvents (water, acetonitrile, methylene chloride, chloroform, and methanol), ammonium acetate (Optima LC/MS), and sodium chloride were purchased from Thermo Fisher Scientific. The following extracts and standards were purchased from Avanti Polar Lipids: lysyl-PG 16:0 (LysylPG 840520), MGDG (Gal₁DG 840523), DGDG (Gal₂DG 840524), CLs (CLs 840012 and 841199), l-α-phosphatidic acid (PA 840101c), and l-α-PG (PG 841138c). The following FFA and diacylglyceride standards were purchased from Nu Chek Prep: pentadecanoic acid (FA 15:0, N-15-A), hexadecanoic acid (FA 16:0, N-16-A), heptadecanoic acid (FA 17:0, N-17-A), octadecanoic acid (FA 18:0, N-18-A), nonadecanoic acid (FA 19:0, N-19-A), eicosanoic acid (FA 20:0, N-20-A), heneicosanoic acid (FA 21:0, N-21-A), 1,3-dimyristoyldiacylglyceride (1,3-DG 14:0, D-142), 1,3-dipalmitoyldiacylglycerol (1,3-DG 16:0, D-152), 1,3-distearoyldiacylglyceride (1,3-DG 18:0, D-162), 1,3-dioleoyldiacylglyceride (1,3-DG 18:1, D-237), cis-1,3-dilinoleoyldiacylglyceride (cis-1,3-DG 18:2, D-252), 1,3-diarachidoyldiacylglyceride (1,3-DG 20:0, D-172), and 1,3-diarachidonoyldiacylglyceride (1,3-DG 20:4, D-297). Stock solutions of lipid standards and standard extracts were prepared at 1 mM in chloroform, from which a 5 µM mixture in 2:1 acetonitrile-methanol was prepared for analysis.