Fig. 2. Dap160 co-localizes with aPKC in neuroblasts. (A-L) Neuroblasts co-stained for Dap160 (A-F) and aPKC (G-L); merged images below. Genotypes, developmental stages and cell cycle stages labeled at top. (B,H) Dap160 colocalizes with aPKC at the apical cortex of mitotic embryonic neuroblasts. (E,K) Endogenous Dap160 is undetectable at the site of aPKC apical cortical localization in larval brain metaphase neuroblasts. (F,L) Overexpression of Dap160 reveals colocalization with aPKC at the apical cortex in larval brain metaphase neuroblasts (90%, n=12). Scale bar: 5 µm. (M) aPKC can immunoprecipitate Dap160 from embryonic lysates. Right lane: aPKC antibody can immunoprecipitate Dap160 (top arrow) and aPKC (bottom arrow). Left lane: the GFP control antibody does not immunoprecipitate either Dap160 or aPKC. (N) Two independently generated Dap160 antibodies detect apical crescents of Dap160 in wild-type neuroblasts or Dap160 overexpression neuroblasts (arrowheads) but not in dap160 mutant neuroblasts (brackets). Roos/Marie antibody from Roos and Kelly (Roos and Kelly, 1998). Scale bar: 5 µm. (O) Dap160 overexpression results in the formation of Dap160/Sec15 double-positive puncta in neuroblasts (arrowheads; left pair of panels) and Dap160/aPKC double-positive puncta (arrowheads; right pair of panels). Scale bar: 5 µm.