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. 2017 Nov 2;8:1462. doi: 10.3389/fimmu.2017.01462

Figure 3.

Figure 3

Nlrp10−/− DCs exhibit impaired IL-12 production after CD40 ligation or CpG DNA stimulation. (A,B) ELISA measurement of IL-12p40 levels in cell supernatants from OT-II CD4+ T cells co-cultured with immature (A) or matured (B) WT or Nlrp10−/− DCs loaded with ovalbumin (20 µg/ml) for 3 days. (C) WT and Nlrp10−/− DCs were incubated with L929 fibroblasts expressing or not expressing CD40 ligand (CD40L). Cell supernatants were collected after 3 days and the IL-12p40 levels were measured by ELISA. (D) ELISA measurement of IL-12p40 production by WT and Nlrp10−/− DCs stimulated by MegaCD40L for the indicated times. (E) WT and Nlrp10−/− DCs were left untreated or stimulated with either CpG DNA or LPS and the IL-12p70 levels in supernatants were then determined at the indicated times. (F) ELISA measurements of IL-12p70 and IL-12p40 production by WT and Nlrp10−/− DCs stimulated for 12 h with increasing concentrations of CpG DNA (0.001–10 µM). (G) Relative expression of Il12a and Il12b in WT and Nlrp10−/− DCs stimulated with CpG DNA (1 µM) for the indicated times and assessed by semi-quantitative RT-PCR. Data shown represent the mean ± SD and represent an experiment performed in triplicates and repeated with at least with three different DC preparations. *p < 0.05; **p < 0.01; ***p < 0.001. Abbreviations: DCs, dendritic cells; LPS, lipopolysaccharide; WT, wild-type.