FIG 5.

IRF9 is important for DC function in response to IFN-I and TLR7 as well as TLR9 ligands. (A to D) WT or Irf9^−/− Flt3L-DCs were left unstimulated (n.c.) or were stimulated with LPS, RNA40 complexed to DOTAP, CpG2216, or IFN-β. (Α) Cytokine concentrations were measured by ELISA after 22 h of stimulation. (B and D) mRNA levels for the indicated genes were analyzed by qRT-PCR analysis and normalized to the levels of Hprt1. Relative expression was calculated by setting the value for unstimulated WT Flt3L-DCs to 1. (C) Immunoblotting of IRF7, MAPK, p65, and β-actin in WT or Irf9^−/− Flt3L-DCs after 22 h of stimulation with RNA40 or CpG2216 or in unstimulated cells (n.c.). For panel A, data were combined from three independent experiments performed in duplicate and are means and SEM. For panels B and D, data were combined from two independent experiments performed in duplicate and are means and SEM. For panel C, data from one of three independent experiments with consistent results are shown. **, P < 0.01; ***, P < 0.001; n.s., not significant (Student's t test).