Influence of extracellular RNA on pro- and anti-inflammatory mediators in bone marrow-derived macrophages (BMDM). (A) mRNA expression of Tnf-α, Arg2, Il-1β, Il-6, Ifn-γ, Il-10, Il-4 and Stat1 in wild-type BMDM, differentiated in the presence of M-CSF-containing L929-conditioned medium was analyzed by real-time PCR in the absence (control, dotted line) or presence of eRNA (1, 10, or 25 µg/ml) for 24 h. Data are expressed as changes in the ratio between target gene expression and Gapdh mRNA. Values represent mean ± SD (n=6 per group); *p<0.05, **p<0.01, ***p<0.001 vs. control, ns = non-significant. (B) mRNA expression of Tnf-α, Arg2, Il-1β, Il-6, Il-12, iNOS, Ifn-γ, Il-10, Il-4, Stat1, Cd206 and Arg1 in wild-type BMDM, differentiated in the presence of mouse recombinant M-CSF was analyzed by real-time PCR in the absence (control, dotted line) or presence of eRNA (1, 10, or 25 µg/ml) for 24 h. Data are expressed as changes in the ratio between target gene expression and Gapdh mRNA. Values represent mean ± SD (n=6 per group); *p<0.05, **p<0.01, ***p<0.001 vs. control, ns = non-significant. (C) TNF-α or (D) IL-6 protein concentration was measured in the supernatants of BMDM following treatment in the absence (control) or presence of eRNA (1, 10, or 25 µg/ml) as indicated. Values correspond to mean ± SD (n=9 per group). *p<0.05, **p<0.01, ***p<0.001, ns=non-significant.