Figure 1. ZNRF1 deletion impairs LPS-induced production of inflammatory mediators in BMDMs.

(a–c) BMDMs were harvested from wild-type (Znrf1^F/F) or Znrf1^Δ mice and treated with LPS (100 ng ml⁻¹) for the indicated times. (a) The expression of the indicated mRNAs was analysed by RT–qPCR. (b) The production of cytokines in supernatants was determined by ELISA. (c) The phosphorylation of MAPKs, IKKα/β, IRF3 as well as the indicated proteins in cell lysates was analysed by immunoblot analysis. The intensities of the bands are expressed as fold increases compared to those of untreated control cells after normalization to their unphosphorylated forms. (d) BMDMs from Znrf1^Δ mice were reconstituted with either Flag-tagged wild-type ZNRF1 or ZNRF1(C184A) mutant and stimulated with LPS (100 ng ml⁻¹) for 8 h. The levels of the indicated cytokines in culture supernatants were measured by ELISA. *P<0.05 (Student's t-test). The data are representative of three independent experiments performed in triplicate (error bars, s.d.).