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Figure 4


Fig. 4. Dap160 positively regulates neuroblast pool size. (A) Neuroblast numbers scored wild-type (green bars), dap160 mutants (red bars) and Dap160 misexpression larvae at second instar (L2) or wandering third instar (L3). Numbers inside bars indicate numbers of brains analyzed. See Materials and methods for genotypes and growth temperatures. (B-E) Wild-type (B,C), dap160 (D) and aPKC (E) mutant clones (n≥50 and n=35 respectively) always contain a single Deadpan-positive neuroblast (arrowhead). (F-L) Neuroblast cortical polarity in larval neuroblasts. (F,G) Wild-type neuroblasts have aPKC apical crescents (arrowhead) and Miranda (Mira) basal crescents. (H,I) dap160 mutants have weak ectopic cortical aPKC (arrows) and normal Mira basal crescents (15%, n=13). (J,K) Dap160 overexpression in neuroblasts leads to weak ectopic cortical aPKC (arrow), increased cytoplasmic and reduced cortical Mira (19%, n=21 as shown; remainder normal basal crescents); Dap160 is detected in cortical patches and cytoplasmic puncta (L). (M,N) Live imaging of wild-type neuroblasts with GFP::Mira and Cherry::Jupiter. (M) Wild-type neuroblasts show basal GFP::Mira at metaphase (brackets) and partition GFP::Mira to the GMC at telophase (arrowhead; 100%, n=37). (N) Dap160 misexpressing neuroblasts show cytoplasmic GFP::Mira at metaphase (brackets) and occasionally do not segregate GFP::Mira to the GMC at telophase (arrow; 8%, n=40). Scale bars: 5 µm.





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