Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA093681

Local Sample ID:	8_2
Subject ID:	SU001361
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090

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Treatment:

Treatment ID:	TR001376
Treatment Summary:	Mouse thioglycollate-elicited macrophages and bone marrow derived macrophages were obtained and cultured as described previously (Heinz et al., 2010). For various ligand treatments, cells were in Phenol-red free RPMI-1640 (Invitrogen) supplemented with either 10% heat-inactivated FBS (Hyclone) or 10% lipid-deficient FBS (Hyclone) for 24hr before treatment, then treated with either 1µM GW3965, 0.25-20µM desmosterol, or 0.3-30µg/ml cholesterol for 12-24hr. 50µM melavonate and 10µM Mevastatin (Sigma) were added to media in desmosterol response experiments. For TLR repression studies, cells were cultured in medium containing Phenol-red free RPMI 1640 (Invitrogen) with either heat-inactivated FBS (Hyclone) or lipid-deficient FBS (Hyclone) for 24hr, then pretreated with either 20-30µM desmosterol, 50µg/ml cholesterol, 5µM triparanol, or 50-100µM 9Z-palmitoleic acid for 18hr. Cells were stimulated with Pam3CSK4 (300ng/ml), PolyI:C (50ng/ml), LPS (100ng/ml), TNFa (30ng/ml), or KLA (100ng/ml) for 6hr. See Extended Experimental Procedures for additional details at https://doi.org/10.1016/j.cell.2012.06.054

Treatment ID:

TR001376

Treatment Summary:

Mouse thioglycollate-elicited macrophages and bone marrow derived macrophages were obtained and cultured as described previously (Heinz et al., 2010). For various ligand treatments, cells were in Phenol-red free RPMI-1640 (Invitrogen) supplemented with either 10% heat-inactivated FBS (Hyclone) or 10% lipid-deficient FBS (Hyclone) for 24hr before treatment, then treated with either 1µM GW3965, 0.25-20µM desmosterol, or 0.3-30µg/ml cholesterol for 12-24hr. 50µM melavonate and 10µM Mevastatin (Sigma) were added to media in desmosterol response experiments. For TLR repression studies, cells were cultured in medium containing Phenol-red free RPMI 1640 (Invitrogen) with either heat-inactivated FBS (Hyclone) or lipid-deficient FBS (Hyclone) for 24hr, then pretreated with either 20-30µM desmosterol, 50µg/ml cholesterol, 5µM triparanol, or 50-100µM 9Z-palmitoleic acid for 18hr. Cells were stimulated with Pam3CSK4 (300ng/ml), PolyI:C (50ng/ml), LPS (100ng/ml), TNFa (30ng/ml), or KLA (100ng/ml) for 6hr. See Extended Experimental Procedures for additional details at https://doi.org/10.1016/j.cell.2012.06.054