Compactin is a potent competitive inhibitor of hydroxymethylglutaryl coenzyme A reductase, the rate-limiting enzyme in cholesterol biosynthesis. Experiments were conducted with peritoneal macrophages, obtained from thioglycolate-stimulated mice, as per protocols PP0000001401, PP0000001502 and PP0000001801.
For the 8 time point study, cells were treated for 0, 0.5, 1, 2, 4, 8, 12, or 24 hours with Kdo2-Lipid A. Untreated cells were used as a zero timepoint. Three separate experiments (biological replicates) were carried out, and a single MS measurement was performed on each sample. Measurements were taken at each timepoint for: (i) Kdo2-Lipid A + compactin, (ii) Kdo2-Lipid A, (iii) compactin, and (iv) control.
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)