Figure 4.

DS activates NF-κB signaling. Activation of NF-κB and STAT signaling was measured by their translocation from endothelial cell cytosol to cell nuclei. (a) Measurement of NF-κB activation by electrophoretic mobility shift assay of nuclear extracts from endothelial cells treated for 0–120 minutes with 50 μg/mL pure DS, or 120 minutes with 2.5 ng/mL TNF-α (lane T). All extracts were mixed with ³²P-labeled oligonucleotides containing the NF-κB binding site. The sample in lane B is identical to that in lane T, with the addition of a 50-fold excess of unlabeled oligonucleotide. (b) Immunostaining of endothelial cells with antibody ISGF3 specific for the STAT complex. (1) Cells treated with media alone for 30 minutes. (2) Cells treated with 50 μg/mL pure DS for 30 minutes. (3) Cells treated with 100 U/mL IFN-γ for 30 minutes.