Figure 4.

A, Coimmunoprecipitation of APP and NEEP21 was performed on brain homogenate from nontransgenic mice. IP with a C-terminal antibody was able to copurify NEEP21. In cell lysates from N2a cells stably expressing APP and NEEP21-myc-his, copurification was observed using both an N-terminal (P2-1) and C-terminal (369) antibody. B, Coexpression of Fe65 with APP and NEEP21 did not increase the efficiency of co-IP relative to coexpression of GFP with APP and NEEP21. C, Using N2a cells transiently expressing NEEP21-myc-his and Fe65, we also failed to observe co-IP, although an APP-Fe65 complex was detectable. Text to the right of each image indicates primary antibody used for Western blot.