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. 2008 Jul 17;27(15):2147–2157. doi: 10.1038/emboj.2008.143

Figure 2.

Figure 2

K7 inhibits TLR-induced NF-κB activation. In (A–C), HEK293 cells were transfected with pRK5-K7R or empty vector and the NF-κB luciferase reporter gene. Cells were stimulated with 20 ng/ml IL-1β for 6 h (A), transfected with 50 ng of CD4TLR4 for 24 h (B) or HEK293-TLR3 cells were stimulated with 25 μg/ml poly(I:C) for 8 h (C). Data are expressed as the mean fold induction±s.d. relative to control levels, for a representative experiment of three, each performed in triplicate. (D, E) For agonist-induced cytokines, HEK-TLR4 (D) or HEK-TLR3 (E) cells were transfected with K7R 24 h prior to stimulation with 1 μg/ml LPS or 25 μg/ml poly(I:C), respectively. After 24 h, supernatants were assayed for IL-8 (D) or RANTES (E) by ELISA. The experiments were performed four times in triplicate and data are expressed as the mean±s.d. from one representative experiment. (F) HEK293T cells were transfected with pCMV-HA-K7R and IRAK2-Myc and 48 h later, lysates were subjected to immunoprecipitation (IP) analysis with the indicated antibodies. (G) HEK293T cells were transfected with pCMV-HA-K7R and Flag–TRAF1, 2, 3, 4, 5 or 6 and 48 h later, lysates were subjected to IP analysis with the indicated antibodies. Results shown are representative of at least three experiments.