DEV00392 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. Cap cells in expanded and ectopic niches express normal cap cell markers. All panels represent one confocal section of a part of the germaria that are labeled for Hts (green, A-C and F), β-gal (red, A-F), Lamin C (green, D,E), DE-cadherin (light blue, F) and DNA (blue, A-E). (A′-F′) are schematics of A-F, in which the areas containing cap cells or ectopic cap cells are highlighted by ovals in A-E and arrowheads in F. (A,A′) A c587-gal4/+; UAS-N^int/wg-lacZ germarial tip showing one β-gal-positive cap cell (arrowhead) among several cap cells and three GSCs (their spectrosomes indicated by arrows) highlighted by an oval. (B,B′) A part of a c587-gal4/+; UAS-N^int/wg-lacZ germarium showing that a group of ectopic cap cells (oval), four of which (two indicated by arrowheads) are β-gal-positive, support seven ectopic GSCs (one of their spectrosomes indicated by an arrow). (C,C′) A part of a c587-gal4/+; UAS-N^int/wg-lacZ germarium highlighted by an oval, showing that a group of cap cells, some of which (two by arrowheads) are β-gal-positive, support eight GSCs (one spectrosome indicated by an arrow), whereas follicle cells are indicated by FC. (D,D′) A wg-lacZ/+ germarial tip showing that one (indicated by an arrowhead) of the Lamin C-positive cap cells (oval) is also β-gal-positive. (E,E′) A part of a c587-gal4/+; UAS-N^int/wg-lacZ germarium showing that a few (one indicated by an arrowhead) of the ectopic Lamin C-positive cap cells (oval) are LacZ-positive. (F,F′) An anterior part of a c587-gal4/+; UAS-N^int/wg-lacZ germarium showing the accumulation of DE-cadherin (several sites indicated by arrowheads) between ectopic cap cells and their associated GSCs. Scale bar: 10 μm.

• Supplemental Figure 2 -

  Fig. S2. Expression of an activated N in inner sheath cells of the adult ovary fail to produce ectopic niches sufficient for maintaining ectopic GSCs. All panels represent one confocal section of a part of the germaria that are labeled for Hts (red, spectrosomes and fusomes), GFP (green, the cells overexpressing dpp in A and an activated N in B and C) and DNA (blue). (A) A hsFLP/+; AyGal4 UAS-GFP/+; UAS-dpp/+ germarium showing that the GFP-positive cells (one indicated by an arrowhead), which ectopically express dpp, can sufficiently support GSCs (one spectrosome indicated by an arrow) located away from the cap cells supporting endogenous GSCs (white arrowhead). (B,C) hsFLP/+; AyGal4 UAS-GFP/+; UAS-N^int/+ germaria showing that the GFP-positive cells (arrowheads), which ectopically express an activated N, cannot sufficiently support GSCs (by presence of branched fusomes indicated by arrows), while the cap cells can support GSCs (filled arrowheads). Scale bar: 10 μm.

• Supplemental Figure 3 -

  Fig. S3. N signaling is required for maintaining the GSC niche in the adult ovary. Panels show one confocal section of the adult germaria that are labeled for β-gal (red, A,B), Hts (green, spectrosomes and fusomes; A-H), Lamin C (red, cap cells; C-H) and DNA (blue, A-H). (A) The Dl-lacZ/+ germarium showing β-gal expression in the two GSCs (solid circles) and the cap cells (indicated by an arrowhead). (B) The E(spl)m7-lacZ/+ germarium showing active Notch signaling in the cap cells (highlighted by broken lines) and no or low Notch activity in the two GSCs (solid circles). (C,D) Germaria from the wild-type females cultured at 29°C for one week (C) and two weeks (D) showing normal cap cells (highlighted by broken lines) and normal numbers of GSCs (three in C and two in D). (E,F) Germaria from the N^ts mutant females cultured at 29°C for one week showing reduced cap cells (highlighted by broken lines) and reduced numbers of GSCs (one in E and none in F). (G,H) Germaria from the N^ts mutant females cultured at 29°C for two weeks showing reduced (highlighted by broken lines in G) or no (H) cap cells and no GSCs (G,H). Scale bar: 10 μm.