Increased expression and activation of IRF-3 in diffDCs. (A) Nuclear extracts from imDCs and diffDCs (1 × 10⁶ cells/mL) cultured in medium alone or stimulated with 500 ng/mL LPS for 10, 20, 40, or 60 minutes were prepared, blotted, and probed with IRF-3–specific antibody. (B) imDCs and diffDCs were cultured in medium alone as control or stimulated with 500 ng/mL LPS for the indicated times. Total cell extracts were probed for IRF-3 and actin as a loading control. (C) Quantification of activated IRF-3 in imDCs and diffDCs after LPS stimulation by assaying IRF-3 DNA-binding activity. Data are shown as means ± SD of 3 independent experiments. *P < .05.