(A—E) Twenty-four hours after injury or sham treatment of BALB/c mice and of MyD88^−/−, TLR4^−/− and RAG2^−/− mice, pLN cells were analysed for the presence of CD3⁻DX5⁺ NK cells and their expression of CD11b. (A) Gating strategy of total CD3⁻DX5⁺ NK cells. The numbers indicate the percentage of NK cells among total pLN cells. (B) Dot plots showing the expression of CD11b on NK cells. The numbers indicate the percentage of CD11b⁺ NK cells among total pLN cells (upper line) and among total NK cells (lower line). Dot plots are representative of three experiments with n=4 mice each. (C) Percentage of CD11b⁺ NK cells among total pLN cells and absolute numbers of CD11b⁺ NK cells per mouse (BALB/c). Data show mean ± S.D. of individual mice from one of the three experiments with n=4 mice each. (D) Percentage of CD11b⁺ NK cells in pLN cells from BALB/c mice (already shown in C) in comparison with pLN cells from MyD88^−/− and TLR4^−/− mice. (E) Percentage of CD11b⁺ NK cells in pLN cells from RAG2^−/− mice. Statistically significant differences were detected with Student's t-test. (F) BALB/c mice were treated with an anti-asialo antiserum (αGM-1) or control serum before and after injury or sham treatment as described in the section ‘Materials and Methods’. OVA-specific T-cells were applied, OVA was injected into the hind footpads and pLN cells were pooled per group (n=3 mice each). The release of IFN-γ from restimulated pLN cells was determined. Data show mean ± S.D. of triplicate cultures and are representative of two experiments. Statistical differences were tested with one-way ANOVA followed by the Bonferroni post-test. *P<0.05; **P<0.01; ***P<0.001.