Fig. 6.

ELF3 induction downstream of IκBζ regulates CCL8 mRNA expression

(A and B) HEK293 cells and OA synovial fibroblasts were transfected with plasmid coding for ELF3 or with empty vector. After 48 h cells were stimulated for 24 h with TNF (10 ng/ml) or left untreated. (A) CCL8 mRNA expression in HEK293 cells and (B) CCL8 mRNA expression in human OA primary synovial fibroblasts. (C) HEK293 cells were transfected with plasmid coding for ELF3. After 48 h dimethyl sulfoxide (DMSO), IMD-0354 (IMD; 2 μM), SP600125 (SP; 10 μM), SB202190 (SB; 10 μM) or U0126 (10 μM) was added for 1 h in the cell culture medium, followed by stimulation with TNF (10 ng/ml) for 8 h. CCL8 mRNA expression was analysed by qRT-PCR. (D) HEK293 cells were transfected with plasmid coding for IκBζ or with empty vector for 24 h, after which siRNA targeting ELF3 or negative control siRNA was introduced to the cells. After 24 h the cells were stimulated with TNF (10 ng/ml) or left untreated. CCL8 mRNA at 24 h after stimulation was analysed by qRT-PCR. Data represent means ± SEM of four (A, C) or three (B, D) individual experiments. *P <  0.05, **P < 0.01, ***P < 0.001. CCL8: chemokine (C-C motif) ligand 8; ELF3: E74-like factor-3; IκBζ: NF-κB inhibitor-ζ.