Figure 6

Segregation of NET and VMAT2 detected via immunoisolation of SCG vesicles. Western blot analysis of immunoisolated membranes captured with NET, VMAT2 or Na/K ATPase antibodies. A. Western blot analysis of NET-05 immunoprecipitates from solubilized mouse cortical synaptosomes (solubilized P2 fraction probed with NET-05 to reveal NET). B. NET immunoreactivity associated with vesicles obtained from lysed mouse cortical synaptosomes from wildtype or NET KO mice, immunoprecipitated with paramagnetic anti IgG beads coated with NET-05 antibody, or beads alone. Panel C shows Western blot analysis of immunoprecipitations of organelles from SCG neuron by beads coated with NET antibodies (organelle isolation, NET beads) and Na/K ATPase antibodies (organelle isolation, Na/K ATPase beads) or VMAT2 antibodies (organelle isolation, control or VMAT2 beads). Elutants of immunoprecpitations are probed with antibodies against either NET or VMAT2.