Figure 3.

Interaction of CB, gephyrin and eIF3 in the brain. Adult mouse whole brain lysates were immunoprecipitated using (A) anti-eIF3H or (B) anti-eIF3B antibodies and endogenous CB (~62 kDa) and gephyrin (~95 kDa) were detected in immunoprecipitated samples using anti-CB and anti-gephyrin specific antibodies respectively. As negative controls, corresponding isotype-matched immunoglobulin G antibodies (including rabbit anti-Sox2 and goat anti-BMP1 antibodies) were used for immunoprecipitaion, and no coimmunoprecipitation of CB and gephyrin was observed. All results shown represent one of at least three independent experiments. Although a faint band corresponding to coimmunoprecipitated CB was detected using anti-eIF3B antibody for immunoprecipitation (B, top left panel), it is worth pointing that the same weak but specific CB band was observed in all the experiments, which suggest that CB also interacts with the eIF3 complex. IP = antibody used for immunoprecipitation; Blot = antibody used for immunoblot analysis; IgG-Hc = immunoglobulin heavy chain (~55 kDa).