rPCN triggers TLR-mediated cell activation.

<p>HEK293T cells were transfected with CD14 and CD36 along with TLR2/1 (A), TLR2/6 (B), or TLR4 (C). The total amount of DNA in each transfection was kept constant by adding empty expression vector. The cells were stimulated with the indicated concentrations of rPCN, previously incubated with polymyxin to neutralize LPS, at 37°C for 20 h. The agonists used as positive controls were: Pam3CysSK4 (P3C) for TLR2/1, fibroblast stimulating ligand-1 (FSL-1) for TLR2/6, and bacterial lipopolysaccharide (LPS) for TLR4. Medium was used as negative control for cell stimulation (white bars). The cell supernatants were analyzed for IL-8 by ELISA. Statistical differences were determined by comparing transfected cells stimulated with rPCN to transfected cells incubated with medium. The results were considered significant when p<0.01 (**) or p<0.001 (***).