Figure 4. During MCMV infection, the TLR9/Myd88‐dependent pathway is necessary for pDC cytokine production, irrespective of their IFN‐I responsiveness.

• A
  Dot plots represent IFN‐I versus GFP staining in 5.2⁺ versus 5.1⁺ pDC isolated from indicated MCMV‐GFP‐infected MBMC. One staining representative of six animals from two independent experiments is shown.
• B, C
  Percentages of GFP⁺ cells within 5.2⁺ pDC (B) or of GFP⁺ (green) versus GFP⁻ (white) cells within 5.2⁺ IFN‐I‐producing pDC (C) isolated from indicated MCMV‐GFP‐infected MBMC. Data shown (mean ± SEM) are from two pooled independent experiments each with three mice per group.
• D
  qPCR for the expression of the indicated genes was performed on whole mRNA isolated from the spleen of indicated MCMV‐infected MBMC. CTR MBMC (black) and Myd88 ^−/− TST MBMC (dark green) were treated with isotype control (IC, square) or blocking α‐IFNAR1 mAbs (triangle). Data shown (mean ± SEM) are from two pooled independent experiments each with two to three mice per group.
• E–G
  CD86 MFI in 5.2⁺ pDC of each indicated MBMC type (E) and 5.2/5.1 ratio of cytokine‐producing pDC (F, G). Data (mean ± SEM) are representative of two pooled independent experiments each with two to three mice per group.
• H–L
  CD86 MFI (H, K) and percentages of cytokine‐producing cells (I, J and L) within pDC of each indicated mouse strain. Black, C57BL6; red, Ifnar1 ^−/−; orange, Ifnar1 ^−/− × Myd88 ^−/−; green, Tlr9 ^−/−; and pale green, Tlr7 ^−/−. In (K–L), indicated mice were treated with IC (square) or blocking α‐IFNAR1 mAbs (triangle). Data depicted (mean ± SEM) are from two pooled independent experiments each with two to three mice per group.

Data information: ns, not significant (P > 0.05); *P < 0.05; **P < 0.01; ***P < 0.001; nonparametric Mann–Whitney test for (B), nonparametric Kruskal–Wallis test combined with Dunn's multiple correction test for (D–L).