Fig. 1.
Partial loss of PntP1 increases the number of type II NBs and eliminates INPs. (A) The generation of Drosophila pntP1-specific mutants. (a) The gene structure of pntP1 and gRNA target sites (arrowheads). Gray boxes, untranslated regions; orange boxes, coding regions. (b) Sequences of gRNA target sites (red) and indels in pntP182 and pntP190 mutants. (c) The protein structure of PntP1 and truncated PntP1 proteins generated from pntP182 and pntP190 mutants. Gray areas, non-specific sequences caused by frame shifts. (B,B′) A wild-type (WT) brain lobe. Type II NB lineages are labeled with mCD8-GFP driven by pntP1-GAL4. Arrows, Ase− type II NBs; arrowheads, Dpn+ Ase+ INPs. (C-F′) pntP1 mutant brains of the indicated genotypes. Brains are stained with phalloidin to outline NBs. The number of Ase− type II NBs (arrows) is significantly increased in pntP190 (C,C′), pntP190/pntΔ88 (E,E′) and pntP182/pntΔ88 (F,F′) mutants but not in pntP182 (D,D′). Dashed circles outline type II NB lineages that generated GMCs (open arrowheads) but not INPs. Insets are enlarged views of lineages without INPs. (G-H′) Pnt knockdown leads to ectopic Ase expression in type II NBs (green arrows) and elimination of INPs in most lineages (dashed lines). Yellow arrows, Ase− NBs; open arrowheads, GMCs. (I-K) Quantification of the number of Ase− type II NBs (I), percentage of Ase− NBs with INPs (J) and number of INPs (K) in pnt mutants. (L-O) Quantification of the total number of type II NBs (L), percentage of lineages with Ase+ NBs (M), percentage of lineages with INPs (N) and number of INPs (O) in Pnt knockdown brains. Sample sizes are indicated above each bar. *P<0.05, **P<0.01. (P) Schematic diagrams of wild-type type II NB lineages and phenotypes in pnt mutant or Pnt knockdown type II NB lineages. Scale bars: 20 μm.