FIG. 1.

In vitro and in vivo interactions of ETO with CoRs. (a) Modular organization of SMRT, N-CoR, and ETO. ID, nuclear receptor interaction domains. TBF-associated factor (TAF)-like and Nervy homology domains of ETO (including Zn fingers) are indicated. (b) Results of a two-hybrid assay using the indicated baits and preys. (c) Pulldown experiments. In vitro-translated SMRT (full-length) and N-CoR (amino acids 1 to 1445) (upper panels) and ETO (lower panel) were precipitated with GST or the indicated GST fusion proteins. (d) Mammalian two-hybrid interaction assay. 293T cells were cotransfected with a VP16ETO expression vector (as indicated) and the appropriate GAL4 fusion protein expression vectors. (e) Coimmunoprecipitations of N-CoR and ETO. Immunoprecipitates with anti-N-CoR antibodies against the N- or C-terminal region and control immunoglobulin G (IgG) were obtained from 293T cells transfected with a Flag-ETO expression vector and blotted with an anti-Flag antibody. The input lane contains 2.5% of the total.