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Fig. 1. Forced expression of an activated N can sufficiently expand niche sizes and generate ectopic niches. (A) Schematic of a crosssection of the anterior part of the germarium. (B,B') A confocal section of a c587-gal4; UAS-GFP female gonad at the larval-pupal transition stage labeled for Hts (red), GFP (green) and DNA (blue), showing that the c587-gal4 driver expresses GFP in most, but not all, somatic cells of the gonad including TFs and IGS precursors. (B') Schematic of the region highlighted by a rectangle in B. Panels C,E,G represent one confocal section of the anterior portion of the germaria labeled for ß-gal (red) and Hts (green), while D,F represent overlayed images. (D'-F') Schematic presentations of the areas highlighted by ovals in D-F, respectively; (G') schematic presentation of G. (C) A hh-lacZ/+ germarial tip showing cap cells (oval) and two GSCs indicated by spectrosomes (arrows). ß-gal-positive TFs are not shown on this confocal section. Arrowhead indicates a branched fusome. (D,D') A c587-gal4/+; UAS-N^int/+; hh-lacZ/+ germarial tip showing over 40 lacZ-positive cap cells at the normal location close to the TF (oval), 15 GSCs evidenced by the presence of spectrosomes (two indicated by arrows) and differentiated germ cell cysts evidenced by the presence of branched fusomes (one indicated by an arrowhead). (E,E') A c587-gal4/+; UAS-N^int/+; hh-lacZ/+ germarial tip showing that a group of 12 lacZ-positive cap cells (oval) located away from the tip support six GSCs (one spectrosome denoted by an arrow and one elongated spectrosome by an arrowhead). (F,F') A c587-gal4/+; UAS-N^int/+; hh-lacZ/+ germarial tip showing that a group of 24 lacZ-positive cap cells (oval) surrounded by follicle cells (FC) support eight GSCs (one spectrosome indicated by an arrow). (G,G') A c587-gal4/+; UAS-N^int/+; hh-lacZ/+ germarial tip showing that lacZ-positive cap cells covering the surface of the anterior half of the germarium support the contacting GSCs indicated by spectrosomes, and a differentiated germ cell cyst evidenced by the presence of a branched fusome (arrowhead). Scale bars: 10 µm. CPC, cap cell; DC, developing cyst; ESC, escort stem cell; FC, follicle cell; FS, fusome; GSC, germline stem cell; IGS, inner germarial sheath cell; IGSP, IGS precursor; SS, spectrosome; TF, terminal filament cell; PGC, primordial germ cell.

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