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. 2013 May 9;9(5):e1003351. doi: 10.1371/journal.ppat.1003351

Figure 2. Nucleotide-binding oligomerization domain (NOD)2-mediated signals induce IL-1β and IL-10 production by peritoneal neutrophils during cecal ligation and puncture (CLP)-induced sepsis.

Figure 2

(A) Serum and peritoneal IL-6, IFN-γ, TNF-α, IL-1β, and IL-10 levels were measured in WT (n = 4) and Nod2−/− (n = 4) mice 24 h after CLP. (B) Peritoneal cells were obtained from WT or Nod2−/− mice 4–6 h after injection with thioglycollate and incubated with or without MDP for 24 h. The IL-1β and IL-10 concentrations in culture supernatants were measured. (C) Serum and peritoneal IL-1β and IL-10 levels were estimated in WT (n = 4) and Nod2−/− (n = 4) mice 4, 12, and 24 h after CLP using ELISA. (D) The NOD2 expression pattern was estimated in sorted F4/80−Ly-6G+ and F4/80+Ly-6G− peritoneal cells of WT mice 4, 12, and 24 h after CLP by real-time PCR. (E) IL-1β and IL-10 transcript levels were evaluated in sorted F4/80−Ly-6G+ and F4/80+Ly-6G− peritoneal cells from WT and Nod2−/− mice 4, 12, and 24 h after CLP. (F) Intracellular IL-1β and IL-10 expression in gated F4/80−Ly-6G+ and F4/80+Ly-6G− peritoneal cells was analyzed by flow cytometry 2, and 12 h after CLP, respectively. (n = 3) *P<0.05, **P<0.01, ***P<0.001 for WT vs. Nod2−/− mice (two-tailed unpaired t-test [a, b], two-way ANOVA [c, e]) Results shown are representative of three independent experiments (mean and SEM).