2.8. Electrophoretic Mobility Shift Assay (EMSA)

The nuclear proteins and ³²P-labeled double-stranded NF-κB oligonucleotide probe were prepared as described previously [28]. Briefly, nuclear proteins were incubated with the probe at room temperature for 20 min, and the probe/protein complex was separated on 6% non-denaturing acrylamide gels. After electrophoresis, the gels were dried by dry-heat vacuum method and the bands were visualized by autoradiography. Sequences of the WT and mutant NF-κB oligonucleotides were 5′-AGTTGAGGGGACTTTCCCAGGC-3′ and 5′-AGTTGAGGGGACTTTCCCAGGC-3′, respectively (the consensus sequence and binding site for NF-κB complexes are underlined).