Experiments were conducted with RAW264.7 cells fed 10% fetal calf serum, as per protocol PP0000001004 (pdf). Measurements were taken at 0, 0.5, 1, 2, 4, 8, 12, and 24hrs for both Kdo2-Lipid A-treated and control cells. Three separate experiments (biological replicates) were carried out, and MS measurements were performed in triplicate (technical replicates).
A Mouse Macrophage Lipidome
Dennis EA, Deems RA, Harkewicz R, Quehenberger O, Brown HA, Milne SB, Myers DS, Glass CK, Hardiman GT, Reichart D, Merrill AH, Sullards MC, Wang E, Murphy RC, Raetz CR, Garrett T, Guan Z, Ryan AC, Russell DW, McDonald JG, Thompson BM, Shaw WA, Sud M, Zhao Y, Gupta S, Maurya MR, Fahy E, Subramaniam S. J Biol Chem
(2010) [Epub ahead of print]
Kdo2-Lipid A of Escherichia coli, a defined endotoxin that activates macrophages via TLR-4.
Raetz C, Garrett T, Reynolds C, Shaw W, Moore J, Smith JD, Ribeiro A, Murphy R, Ulevitch R, Fearns C, Reichart D, Glass C, Benner C, Subramaniam S, Harkewicz R, Bowers-Gentry R, Buczynski M, Cooper J, Deems R and Dennis E. Journal of Lipid Research 47, 1097-1111 (2006)