FIG. 8.
LXXLL motif-containing cofactors interact with both ERα and ERβ in vitro in a ligand-dependent manner. Equal amounts of full-length ERα, ERβ, or control BSA were immobilized on 96-well plates in the presence or absence of 1 μM estradiol. Full-length RIP140, GRIP-1, and TRAP220 were translated in vitro and labeled with [35S]methionine. Labeled cofactors were added to the wells containing immobilized protein and incubated at 4°C overnight. Unbound protein was removed by washing, and the bound protein was eluted, separated by SDS-PAGE, and visualized by autoradiography.