Figure 4.

Nucleosome-induced CD69 expression is increased in spleen cells of DNase1-deficient mice. Spleen cells from wild type (WT), DNase1-deficient mice (DNase1^−/−/Trap1^m/m) and TLR2/TLR4-deficient mice were cultured in the presence/absence of different concentrations of purified nucleosomes (Nuc), the nucleosome purification buffer (gradient), LPS (TLR4 agonist) or CpG-ODN (TLR9 agonist). CD69 expression was estimated by flow cytometry. (A) Shown is one representative experiment of eight independent experiments using four independent chromatin preparations. Means and SD of triplicates are depicted. (B) Data pooled from the eight independent experiments with eight mice/group are shown. Means and SEM of individual means are depicted. *p ≤ 0.05; **p ≤ 0.01 (two-tailed paired t-test or two-tailed Wilcoxon matched-pairs signed rank test). (C) Shown is one representative experiment of two independent experiments. Means and SD of triplicates are depicted. (D) Shown is one representative experiment of three independent experiments. Means and SD of triplicates are depicted. MFI, mean fluorescence intensity. Cells were gated on total living cells according to size and granularity and exclusion of dead (propidium iodide-positive) cells.