Fig 5.

TLR2-mediated stimulation levels by lipid 654 and lipid 430. HEK293 cells, transfected with the human TLR2, MD-2, CD14, and SEAP genes, were used to assay the function of lipid 654 and lipid 430 in vitro. For antibody blocking, cells were preincubated for 1 h with neutralizing anti-human TLR2 antibody (10 μg/ml; InvivoGen). HEK293 cells were stimulated for 24 h with DMSO (vehicle; 50% mixture of DMSO-water; approximately 1.11% DMSO in final culture medium; n = 4), the known TLR2 ligand MMP (0.1 μg/ml; n = 6), the known TLR2 ligand LTA (1 μg/ml; n = 6), lipid 430 (0.69 μg/ml; n = 2), and lipid 654 (0.69 μg/ml; n = 7). The sample sizes (n) for each treatment refer to the number of both untreated and TLR2-blocked samples. Responses were assessed 24 h after the addition of TLR2 ligands, and results are expressed as the ratio of stimulated to nonstimulated (DMSO) responses.