Upregulation of TLR1, TLR2, TLR4, and IRAK-2 Expression During ML-1 Cell Differentiation to Macrophages: Role in the Potentiation of Cellular Responses to LPS and LTA

Figure 5

Enhanced LPS and LTA responsiveness in ML-1 cells. One millions cells were incubated for 3 days with medium alone or medium containing 5 nM TPA. Cells were washed after 3 days with PBS to remove TPA, followed by incubation with medium alone for additional three days to complete the differentiation process. Six days differentiated cells were gently washed with PBS and incubated in the absence or in the presence of 100 ng/mL LPS or 10 ng/mL LTA for 16 h. The culture medium was collected, and the levels of selected inflammatory mediators proteins (IL-1 a, IL-1 b, IL-6, IL-8, IL-10, IL-12(p40), GMCSF, IFN-gamma, TNF-a, and IP-10) in cell culture medium were determined using cytokine multiplex immunoassay reagents (Upstate Biotechnology, Lake Placid, New York, United States) analyzed by Luminex 100 (Luminex Corporation, Austin, TX, USA). Data are mean −SE of four separate experiments.