Fig. 3.

IκBζ is synergistically induced by co-stimulation with IL-17A and TNF

(A) Time course of IκBζ mRNA expression in OA synovial fibroblasts stimulated with IL-17A (50 ng/ml) and/or TNF (10 ng/ml). (B) IκBζ immunofluorescence of OA synovial fibroblasts stimulated for 3 h with IL-17A (50 ng/ml) and/or TNF (10 ng/ml). (C) OA synovial fibroblasts were pretreated with cycloheximide (CHX; 5 μg/ml) or dimethyl sulfoxide (vehicle) for 1 h followed by stimulation with IL-17A (50 ng/ml) and TNF (10 ng/ml) for 1–3 h. IκBζ mRNA expression was analysed by qRT-PCR. (D) OA synovial fibroblasts were stimulated with IL-17A (50 ng/ml) and/or TNF (10 ng/ml) for 40 min, after which actinomycin D (ActD; 5 μg/ml) was added. IκBζ mRNA remaining at each time point after addition of actinomycin D was analysed by qRT-PCR. Data represent means ± SEM of three (A, C, D) individual experiments with cells from different donors. *P < 0.05, **P < 0.01, ***P < 0.001 for indicated comparisons. #P < 0.05 compared to control. DAPI: 4′,6-diamidino-2-phenylindole; IκBζ: NF-κB inhibitor-ζ.