Figure 3.

Heparan sulphate (HS) -induced nuclear factor-κB (NF-κB) activation in RAW264.7 cells was inhibited by paxilline. RAW264.7 cells were incubated for 1 hr at 37° with HS (1 μg/ml) in the absence and presence of paxilline (2 μg/ml), respectively. The nuclear proteins were extracted from macrophages and incubated with 1 μl of ³²P-labelled oligonucleotide for 20 min. Binding reactions were then performed in 1 × binding buffer and 0·05 mg/ml poly(dI-dC) in a total volume of 10 μl. Then, 1 μl of loading buffer was added and the reactions were loaded on a 4% non-denaturing polyacrylamide gel. After electrophoresis the gels were dried for autoradiography. Cells treated with lipopolysaccharide (LPS; 10 ng/ml) in the absence and presence of paxilline, respectively, served as control. *P < 0·05, HS + paxilline versus HS; ^#P < 0·05, LPS + paxilline versus LPS. Results of one out two experiments with similar results are shown.