Figure 4.
BLOC-1 interacts biochemically and genetically with the AP-3 complex. A, B, Crosslinked lysates from HEK and SH-SY5Y cells stably transfected with FLAG-dysbindin were immunoprecipitated with the FLAG antibody (lanes 2, 2′). Lanes 1, 1′ contain the total cellular lysate input and as a negative control the immunoprecipitation was done in the presence of the antigenic peptide (out competition control; lanes 3, 3′). Samples were analyzed by immunoblotting with BLOC-1 subunit antibodies and AP-3 subunit antibodies. C–J′, Dentate gyri from wild-type (C, E, G, I, K), AP-3-null (Ap3d1mh/mh, D, H), and BLOC-1-null mouse brains (Pldnpa/pa, M; Mutedmu/mu, J; and Dtnbp1sdy/sdy, L) were stained with antibodies against AP-3 δ and VAMP2 as a control. C′–M′ depict AP-3 δ staining; note the marked alterations in the AP-3 δ staining in the AP-3-null Ap3d1mh/mh brains.