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. 2013 Oct 3;140(3):314–322. doi: 10.1111/imm.12139

Figure 3.

Figure 3

Knockdown of nuclear factor-κB (NF-κB) repressing factor (NKRF) inhibits lipopolysaccharide (LPS) -induced NKRF expression, NF-κB activity, the expression of cytochrome oxidase subunit 2 (COX-2) and interleukin-6 (IL-6). (a, b) RAW264.7 cells were transfected with si-NKRF or control small interfering (si) RNA at a final concentration of 100 nmol/l (M). After 48 hr, the cells were stimulated with LPS for 6 hr, and then NKRF expression was detected by quantitative PCR (qPCR) (a) and Western blot assay (b). (c) RAW264.7 cells stably transfected with pGL4.32[luc2P/NF-κB-RE/Hygro] were transfected with si-NKRF for 48 hr. After the cells were stimulated with LPS (100 ng/ml) for 6 hr, firefly luciferase activity was measured and was normalised to Renilla luciferase activity. (d, e) The mRNA and protein levels of COX-2 were determined by qPCR and Western blot. (f, g) The mRNA and protein levels of IL-6 were analysed by qPCR (f) and ELISA (g) in LPS-treated macrophages. Data are presented as the mean ± SD (n = 3) of three independent experiments. *P < 0·05; **P < 0·01.