Transcriptomic profile of trametinib on LPS-mediated gene expression in murine peritoneal macrophages

To determine the underlying mechanisms by which trametinib affected LPS-induced inflammatory response in macrophages, we performed microarrays to define the global gene expression in murine peritoneal macrophages treating with trametinib or vehicle followed by LPS stimulation. Overall design: We injected three mice with thioglycollate broth intraperitoneally. Three days later, the mice were injected with 5 ml of cold PBS into the peritoneal cavity and gently massaged. Then, the peritoneal fluid was collected, centrifuged, and the cell pellet was resuspended in DMEM supplemented with 10% FBS and cultured in 6-well-plate at 37?. After adhering for 1h, the cells were gently washed with warm PBS for 3 times to remove non-adherent cells and then cultured with fresh medium. After additional 24h of culture, these cells were treated with trametinib (100nM) or vehicle (DMSO) for 2h, and then challenged by LPS (100ng/ml) for 12h.