Figure 4.

DRM- and DSM-associated IDE pools are proteolytically active in vitro. A- Western blotting with BC2 anti-IDE polyclonal antibody of immunoprecipated IDE from isolated DRM and DSM fractions of rat brain using 1C1/3A2 anti-IDE monoclonal antibodies. B- Representative phosphorimage scan showed degradation of [¹²⁵I]-insulin and [¹²⁵I]-Aβ after incubation with anti-IDE immunoprecipitates from DRM and DSM in the presence of a protease inhibitor cocktail (as defined in Antibodies and Chemicals) with or without metalloprotease inhibitors (EDTA/1,10 phenantroline). IgG, unrelated immunoglobulin used as a negative control for the immunoprecipitation. The intensity of the band in the presence of unrelated IgG and after incubation in degradation buffer was referred as intact substrate (0% degradation). C- Bars represent the semi-quantitative analysis of the percentage of [¹²⁵I]-insulin and [¹²⁵I]-Aβ degradation by IDE from DRMs and DSMs in the presence and absence of 1,10-Phe/EDTA (n = 3; *p < 0.001).