Figure 5: CARD9/BCL10 selectively controls NF-κB activation required for pro-IL-1α/β synthesis.

(a) Pro-IL-1α and pro-IL-1β levels in BMDCs from indicated mice stimulated with TNBS (100 μg ml⁻¹) or LPS (1 ng ml⁻¹) for 12 h in the presence of z-VAD-fmk (100 μM). Cells were lysed in hypotonic buffer and intracellular pro-IL-1α and pro-IL-1β levels were determined by ELISA (means±s.d., n=3 per group). (b) Immunoblotting of NF-κB, c-Rel, p65 and lamin B in the nuclear extracts of WT and CARD9^−/− BMDCs stimulated with LPS (10 ng ml⁻¹), ox-zymosan (100 μg ml⁻¹) or TNBS (100 μg ml⁻¹). The nuclear protein lamin B served as a loading control. Representative data of three independent experiments were shown. (c) IL-1β secretion by WT, CARD9^−/− and BCL10^−/− BMDCs stimulated with LPS (1 ng ml⁻¹) or TNBS (100 μg ml⁻¹) or with TNBS after LPS priming. DAP12-CARD9 signalling pathway is crucial for cytokine response of DCs to TNBS (means±s.d., n=3 per group). (d–f) Production of TNF-α (d), IL-12p70 (e) and IL-6 (f) by WT, MyD88^−/−, CARD9^−/−, FcRγ^−/− or DAP12^−/− BMDCs after stimulation with LPS (10 ng ml⁻¹), Pam3CSK4 (100 ng ml⁻¹), Ox-zymosan (100 μg ml⁻¹) or TNBS (100 μg ml⁻¹) for 24 h (means±s.d., n=3 per group).