TEL and TEL-AML1 interact with N-CoR in vitro.
The in vitro–translated TEL, TEL(Δ53-116), TEL(Δ119-336), TEL-AML1, TEL(Δ53-116)-AML1, and/or AML1 proteins were evaluated for their abilities to interact with (A, B)35S-methionine–labeled N-CoR or (C) SMRT as well as with (D-F) the indicated amino- and carboxy-terminal deletions of N-CoR. The co-immunoprecipitation with mSin3A shown in panel F, lanes 5 and 6, was used as a positive control. The numbers represent the first and the last amino acid in a given protein (or its deletion mutant). In panels A-F, lane 1, 20% of the input is shown. Specificity of the antibody used for a given co-immunoprecipitation is indicated above each panel. In the absence of a given antibody target protein (TEL or AML1), neither full-length N-CoR nor its deletion mutants (amino acids 1-1461, 1586-2453, or 1-758) were co-immunoprecipitated by anti-TEL (shown in panels A, B, and F, lane 2, and panels D and E, lanes 4 and 5, respectively) or anti-AML1 (shown in panel A, lane 3). Prior to their use in a given co-immunoprecipitation reaction, levels of each in vitro translated protein were evaluated by Western blotting to ensure that approximately equal amounts of all antibody-specific input proteins were used for each experiment (data not shown).
