Fig. 5.

Transmembrane tumour necrosis factor (TNF) signalling via TNF receptor 1 (TNFR1) in interferon (IFN)-γ-induced macrophages is sufficient to induce nitric oxide. BMDMϕ from wild-type mice were incubated with 0 U/ml (media alone), 20 U/ml or 100 U/ml IFN-γ for 24 h in the presence of either 10 µg/ml human IgG or sTNFR-Ig (a) or 10 µg/ml I-XPro or XPro1595 (b). BMDMϕ from wild-type and TNFR1^−/− were also incubated with 0–100 U/ml IFN-γ in the presence of XPro1595 (all samples, c). Nitric oxide in the supernatant was quantified. Bone marrow-derived macrophages (BMDMϕ) from wild-type mice were also incubated with media alone or 100 ng/ml lipopolysaccharide (LPS) for 24 h in the presence of 10 µg/ml immunoglobulin (Ig)G or soluble TNFR-Ig (sTNFR-Ig) (d) or 10 µg/ml I-XPro or XPro1595 (e). Mean ± standard error shown; n = 3–4.