Figure 5.
CO inhibits activation of NF-κB and nuclear translocation of IRF-3 by LPS, but not by poly(I:C). RAW 264.7 cells were stimulated with 100 ng/ml LPS, 100 ng/ml Pam, 10 ng/ml Fla, 1 μM CpG, and 20 μg/ml poly(I:C) in the presence or absence of CO. Nuclear protein was extracted for NF-κB activation assay or Western blot analysis. (A) NF-κB (p65) activation was analyzed by ELISA-based kits. **, P < 0.01 versus CO-treated cells. (B) NF-κB was analyzed by electrophoretic mobility shift assay. Protein–DNA complexes are shown. Arrowhead indicates nonspecific bands. (C) Nuclear extract was analyzed by immunoblotting for IRF-3 nuclear translocation, followed by stripping and reprobing for upstream stimulatory factor 2 (USF2).