Figure 2

Electrophoretic Mobility Shift Assay (EMSA) EMSA were performed with 10 μg whole cell extracts and a 10 fmol [gamma-³²P] labeled probe. The specificity of binding to the respective probes was determined by using a 200-fold molar excess of unlabeled oligonucleotide as a competitor, which completely removed all the binding, indicating that these bound complexes resulted from sequence specific DNA-protein interactions. The arrows indicate the LXRα / RXRα complexes (i) and supershifted complexes (ii), respectively. Lanes: 1, LXRα / RXRα; 2, with the addition of unlabeled DR4 oligonucleotide; 3, with the addition of unlabeled mutated LXRE oligonucleotide; 4, supershifted complex; 5, with the addition of unlabeled DR4 oligonucleotide; 6, with the addition of unlabeled mutated LXRE oligonucleotide.