Fig. 5.

N-Myc directly interacts with Sp1 and HDAC1 through its carboxyl-terminal domain. (a) Protein coimmunoprecipitation (IP) of N-Myc or HDAC1. One milligram of nuclear protein extract from LAN-1 cells was incubated with either a preimmune serum, or an anti-N-Myc antibody (Left) or an anti-HDAC1 antibody (Right). The purified IP-complex was analyzed by Western blot, using antibodies for the following proteins: Sp1, HDAC1, Max, and Tip-60. Lane 1, input; lane 2, preimmune serum IgG IP; lane 3, anti-N-Myc or anti-HDAC1 antibody IP. (b) GST-N-Myc fusion proteins carrying different N-Myc domains were incubated with nuclear extracts expressing HA-HDAC1 or HA-Tip60. GST pull down complexes were analyzed by Western blot analysis, using an anti-HA monoclonal antibody.