FIGURE 5.

Association of NF-κB to CXCR3 ligand chemokine promoters. BM-MSCs were either untreated or treated with 10 ng/ml TNFα for 4 h, and nuclear extracts were prepared. EMSA was done with ³²P-labeled oligonucleotide probes corresponding to positions −172 to −135 of the CXCL9 promoter containing wild-type (WT κB) or mutant NF-κB sites (mt κB) (A), −182 to −161 (left) or −128/−106 (33) of the CXCL10 promoter containing wild-type (WT κB) or mutant NF-κB sites (mt κB) (B), or −70 to −51 of the CXCL11 promoter containing wild-type NF-κB site (WT κB) (C). For the competition assay, unlabeled oligonucleotides (Competitor) were added at 10- or 100-fold molar excess before the addition of labeled probes. The supershift assay was performed by the addition of an anti-NFκB (p65) antibody. Arrows indicate DNA-NF-κB complexes. Arrowheads indicate supershifted DNA-NF-κB complexes.