BMMФ were stimulated either with IL-4 (10 ng/ml, AF-214-14, Peprotech), lipopolysaccharide (100 ng/ml, Sigma) or dexamethasone (100nM, D4902, Sigma) to respectively achieve M(IL-4), M(LPS) or M(GC) polarization state. In some experiments, BMMФ were treated with C75 (Sigma), Cerulenin (Sigma), Etomoxir (Sigma), pan-AKT inhibitor MK-2206 (1 mM, Cayman Chemical), 25-hydroxycholesterol (10 μM, Cayman chemical), N-acetyl cysteine (10mM, Sigma), Diphenyleneiodonium chloride (DPI, 5 μM, Sigma), L-NG-Nitroarginine methyl ester (L-NAME, 1mM, Cayman Chemical), Allopurinol (100 μM, Sigma), hydrogen peroxide (Sigma), SIRT1 activator II (Sigma), EX-527 (Sigma), Compound C (Sigma), AICAR (Abcam), HMGCoA (Sigma), water-soluble cholesterol (Cholesterol–methyl-β-cyclodextrin, Sigma) or Simvastatin (Sigma).
For fatty acid treatment, palmitic or oleic acid (Cayman) were solubilized at 100mM in absolute ethanol at 60°C. Fatty acid were conjugated at the desired concentration using a sonicator water bath into the macrophage culture medium to avoid endotoxin contamination from BSA54 (link).
For fatty acid treatment, palmitic or oleic acid (Cayman) were solubilized at 100mM in absolute ethanol at 60°C. Fatty acid were conjugated at the desired concentration using a sonicator water bath into the macrophage culture medium to avoid endotoxin contamination from BSA54 (link).
