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Fig. 2. Ectopic GSCs associated with ectopic niches behave like normal GSCs. The panels A-C and E-H represent one confocal section, whereas D represents an overlayed image of multiple confocal sections. The germaria in A-E are labeled for GFP (green), Hts (red) and DNA (blue), while the ones in F-H are labeled for ß-gal (red), Hts (green) and DNA (blue). (A) A wild-type germarial tip showing that two GSCs (white arrowheads) in contact with cap cells (oval) do not express bam-GFP (black arrowhead). (B) Part of a c587-gal4/+; UAS-N^int/+; bam-GFP/+ germarium showing that two spectrosome-containing GSCs (white arrowheads) that are associated with ectopic cap cells (oval) fail to express bam-GFP. Black arrowheads indicate branched fusomes in differentiated cysts. (C) A middle portion of a c587-gal4/+; UAS-N^int/+; bam-GFP/+ germarium showing that ectopic cap cells (oval) support one bam-GFP-negative spectrosome-containing GSC (arrowhead). (D) The tip of a c587-gal4/+; UAS-N^int/+; bam-GFP/+ germarium showing that the expanded cap cells (indicated by unbroken outline) repress bam expression not only in GSCs (white arrowheads) but also in the spectrosome-containing single germ cells lying more than one cell diameter away (black arrowheads). (E) c587-gal4/+; UAS-N^int/+; bam-GFP/+ germarium showing that the ectopic cap cells (indicated by unbroken outline) repress bam expression not only in GSCs (white arrowheads) but also in the spectrosome-containing single germ cells lying more than one cell diameter away (black arrowhead). (F) The tip of a Dad-lacZ/+ germarium showing that two GSCs (arrowheads) are in contact with cap cells (oval) and express high levels of Dad-lacZ. (G) The tip of a c587-gal4/+; UAS-N^int/+; Dad-lacZ/+ germarium showing that an increased number of cap cells (indicated by unbroken outline) induced by the expression of an activated N support an increased number of GSCs (arrowheads) and cystoblasts that are positive for Dad-lacZ. (H) A middle portion of a c587-gal4/+; UAS-N^int/+; Dad-lacZ/+germarium showing that a group of ectopic cap cells (indicated by unbroken outline) close to follicle cells also support ectopic GSCs (arrowheads) that are also positive for Dad-lacZ. Scale bar: in A, 10 µm for all images.

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