Fig. 2. The NRAGE MHD domain is required for Che-1 binding. (A) Schematic representations of both Flag-tagged NRAGE full-length and its derived deletion mutants. Construct names and binding activities are indicated on the left. (B) Cell lysates from NIH3T3 cells co-transfected with expression vectors encoding the indicated Flag-NRAGE proteins and Myc-Che-1 were immunoprecipitated with anti-Flag antibody. Immunoprecipitated samples were immunoblotted with anti-Myc monoclonal antibody to identify NRAGE domains involved in Che-1 interaction (top) and with anti-Flag antibody for the presence of Flag-NRAGE deletion mutants. Note that, the Flag-NRAGE-g mutant molecule was not revealed in the blot shown here, but under different – more appropriate – gel condition (data not shown).