Fig. 1.

The cholesterol 25-hydroxylase gene is induced by LPS. Bone marrow-derived macrophages were differentiated in vitro with GM-CSF for 8 d and then treated with 10 ng/ml repurified E.coli LPS or media only for 2 h (A). Total RNA was harvested, purified, and labeled, and gene expression analyzed by microarrays (Operon) with oligonucleotide reporters representing approximately 13,000 genes. Cells from four individual mice were used and analyzed in duplicate (n = 8 microarrays total). In B, macrophages were treated with repurified E. coli LPS, live E. coli (one bacterium per macrophage), or media only for 0, 0.5, 1, 2, 4, 6, 12, and 24 h as indicated. Each line represents macrophages taken from one individual mouse. In C, macrophages were derived from Tlr4^−/−, Myd88^−/−, or C57Bl/6 control mouse bone marrow and treated with 10 ng/ml repurified E. coli LPS or media only for 2 h. Cells from four individual mice were used and analyzed in duplicate (n = 8 microarrays total). Fold changes represent expression compared with media controls, i.e., a fold change of 1 indicates no change from media only treated cells. Average data ± SD are presented. Statistics were calculated with a Mann-Whitney test.