OxPAPC does not inhibit NF-κB activation. (A) Immature DCs or DCs differentiated in the presence of 10 (Ox 10) or 25 (Ox 25) μg/mL OxPAPC were stimulated with LPS or PGN. After 70 minutes, total nucleoprotein was extracted. 32P-labeled oligonucleotides containing a NF-κB consensus sequence were incubated at room temperature with 5 μg of nuclear extracts, followed by nondenaturating gel electrophoresis. Similar results were obtained in 2 independent experiments. (B) HEK 293 cells were transfected with a constitutively active TLR4 and a 5xNF-κB firefly luciferase reporter construct. A Renilla reporter plasmid was cotransfected. Firefly luciferase activity with or without addition of OxPAPC (50 μg/mL) was determined after the indicated time points and normalized to Renilla luciferase. Data are presented as mean values ± SD of 3 independent experiments.
