Fig. 5: CLEC2D/TLR2 heterodimers and CLEC2D homodimers negatively regulate IRF5-mediated IL-12 production. | Nature Communications

Fig. 5: CLEC2D/TLR2 heterodimers and CLEC2D homodimers negatively regulate IRF5-mediated IL-12 production.

From: C-type lectin receptor 2d forms homodimers and heterodimers with TLR2 to negatively regulate IRF5-mediated antifungal immunity

Fig. 5

a KEGG analysis of differentially expressed genes involved in signaling transduction in wild-type or Clec2d-deficient BMDCs stimulated with curdlan (20μg/well) for 3 h. b Heatmaps show of differentially expressed genes encoding proinflammatory cytokines in wild-type or Clec2d-deficient BMDCs stimulated with curdlan (20μg/well) for 3 h. c Real-time quantitative PCR analysis of Il12a and Il12b mRNA expression in wild-type and Clec2d-deficient BMDCs stimulated with curdlan (20 μg/well) for 3 h. d Real-time quantitative PCR analysis of Il12a and Il12b mRNA expression in RAW264.7 cells stably expressing mouse Clec2d or mock stimulated with curdlan (20 μg/well) for 3 h. e ELISA results of IL12p70 and IL-12p40 in the supernatants of wild-type and Clec2d-deficient BMDCs, which were stimulated with curdlan (5 μg/well), IFN-γ(20 ng/well), curdlan plus IFN-γ for 6 or 24 h. f Chromatin immunoprecipitation (with control IgG, anti-IRF5 or anti-H3 as positive control; horizontal axis) and PCR analysis of the binding of IRF5 to the promoters of il12a and il12b in wild-type and Clec2d-deficient BMDCs stimulated with curdlan (20 μg/well) for 3 h. g, h ELISA of IL12p70 in the supernatants of wild-type and Clec2d-deficient BMDCs, which were pretreated with IRF5 inhibitory peptide (N5-1, 50 μM) and control peptide (PTD, 50 μM) (g) or MyD88 inhibitor (TJ-M2010-5, 40 μM) (h) and then stimulated with curdlan plus IFN-γ for 6 h. i ELISA of IL-12p70 and IL-12p40 in the supernatants of wild-type, Clec2d-deficient, TLR2-deficient and Clec2d/TLR2-deficient BMDCs stimulated with curdlan or Pam3CSK4 (100 ng/ml) plus IFN-γ (20 ng/well) for 6 h. ns, no significance. Data were presented as mean ± SEM; n = 3 (a, b, d, g Unsti group, h, i), n = 4 (c, e, f, g Curdlan + IFN-γ group) biologically independent samples. Data were analyzed by one-way ANOVA adjusted for multiple comparisons in (a–i). Source data are provided as a Source Data file.

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