Impaired interaction of IRF-3 and CBP in neonatal moDCs. (A-B) Analysis of IRF-3 and NF-κB p65 translocation in adult and cord blood moDCs. (A) Cells were incubated in the presence of medium (unstimulated) or LPS (100 ng/mL) for 2 hours, fixed, and stained for IRF-3 (Alexa 488, green), p65/RelA (Alexa 568, red). Representative fields of IRF-3 and p65 fluorescence from 5 adults and 6 neonates are shown. (B) Nuclear translocation of IRF-3 and p65 were analyzed by Western blotting as described in “Material and methods.” One representative adult and cord blood sample of 3 are shown. (C) Impaired interaction of IRF-3 with CBP in LPS-stimulated cord blood moDCs. Coimmunoprecipitation experiments were performed as described in “Material and methods.” The asterisk (*) denotes IgG. Two representative adult and cord blood samples of 6 are shown. (D) Assessment of IRF-3 DNA-binding capacity in adult and cord blood moDCs. Cells were left untreated or stimulated for 2 hours with LPS. Nuclear extracts were isolated, and IRF-3 DNA-binding activity was measured using TransAM transcription factor assay kit. Results are presented as mean and SEM from at least 5 different donors. *P <.05 as compared with adult moDC.