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. 2017 Aug 22;8:317. doi: 10.1038/s41467-017-00384-z

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — A RNA interference screen for factors mediating glial branch morphogenesis. a Schematic representation of screen strategy. To knockdown genes that encode selected secreted and cell surface molecules, UAS-RNAi and UAS-dcr2 were expressed in astrocyte-like glia under the control of loco ^1.3D2 -Gal4. A secondary screen was performed for UAS-RNAi lines causing early lethality. To achieve mosaic expression of UAS-RNAi transgenes in astrocyte-like glia using loco ^1.3D2 -Gal4, a heat shock-FLP (hs-FLP) recombinase transgene was induced transiently to promote the random excision of the FRT site flanked Gal4-repressor Gal80 encoding cDNA downstream of the widely active tubulin enhancer (tub > Gal80>)⁶⁸. b Lapsyn encodes a transmembrane (TM) cell adhesion protein containing seven leucine-rich repeat (LRR) domains flanked by N-terminal LRR-NT and C-terminal LRR-CT1domains. SP signal peptide. c–e loco ^1.3D2 -Gal4 drives expression of UAS-cd8GFP, UAS-dcr2, and two independent RNAi constructs, UAS-lapsyn ^IR-KK102333 and UAS-lapsyn ^IR-15658-R1. f, g loco ^1.3D2 -Gal4 drives expression of UAS-FB1.1 (white) in controls (f) and UAS-lapsyn ^IR-KK102333, UAS-dcr2 in lapsyn knockdown animals (g). La lamina. Compared to controls (c, f), processes of astrocyte-like mng are reduced following lapsyn knockdown (d, e, g) in the medulla (Me) (d, e, arrowheads), lobula (Lo), and lobula plate (Lop) (d, e, arrows). Some brains displayed a medulla rotation defect (e, asterisk). Panels c–e show single optical sections, f, g projections. For genotypes and sample numbers, see Supplementary Table 1. Scale bars, 50 μm (c–e) and 10 μm (f, g)