Figure 6

Combined effects of LPS and bacterial DNA on pro-inflammatory cytokine production Panels A-C. RAW264 cells were stimulated for 24 h with medium control, LPS [100 ng/ml], stimulatory CpG oligonucleotide AO-1 (3 μM), non-stimulatory GpC oligonucleotide NAO-1 (3 μM), E. coli DNA (bDNA, 5 μg/ml), DNase I-treated bDNA (5 μg/ml) or combinations as indicated. Cell supernatants were harvested and IL-6 (Panel A), IL-12 (Panel B] and TNF-β (Panel C) levels were estimated by ELISA. Data are mean of triplicates ± SD and similar results were obtained in 3 independent experiments. Panel D. Bone marrow-derived primary macrophages were stimulated with medium control, LPS (100 ng/ml), AO-1 (3 μM), NAO-1 (3 μM), E. coli DNA (bDNA, 5 μg/ml), DNase I-treated E. coli DNA (5 μg/ml), or combinations as indicated. After 24 h, cell supernatants were harvested and IL-12 levels in supernatants were estimated by ELISA. Data (mean of triplicates ± SD) are representative of 4 independent experiments.