Figure 3.

Defect of type I IFN production by IRF8^R294C pDCs is not due to increased cell death. FLDC cultures from IRF8^WT and IRF8^R294C mice were kept unstimulated or stimulated with CpG and stained for different surface markers followed by 7-AAD staining. Cells with negative staining for 7-AAD were considered as live cells. Flow cytometric analysis (A) and quantitation (B) of live CD11c⁺ B220⁺ SiglecH⁺ pDCs from IRF8^WT and IRF8^R294C FLDC cultures after 6 h time point. Flow cytometric analysis (C) and quantitation (D) of live CD11c⁺ B220⁺ SiglecH⁺ pDCs from IRF8^WT and IRF8^R294C FLDC cultures after 24 h time point. IRF8^WT activates expression of luciferase reporter construct driven by Ifnb (E) and Ifna6 (F) promoters, while IRF8^R294C was unable to induce the expression of promoters. Data (A, C) are representative of three independent experiments. Data (B, D, E, F) are mean of three independent experiments with error bar representing + SEM and ***p < 0.001, ns, non-significant. p value obtained from Student’s t test.