FIGURE 8.

EMSA experiments show that wild-type FLRev-erbβ and H568F and H568F/C384A variants bind DNA and MBP-NCoR1–540 with similar affinity. Binding assays were prepared in 0.5× TNGD buffer containing 25 ng μl⁻¹ poly(dI-dC), 1 μm FAM-Rev-DR2 (A), or FAM-Bmal1 promoter (B) and the indicated concentrations of wild-type FLRev-erbβ_MGC, or variant FLRev-erbβ, and MBP-NCoR1–540. Assays were incubated on ice for 2 h prior to separation by native gradient gel electrophoresis and imaging of the protein-DNA complexes.