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. 2020 May 8;11:2300. doi: 10.1038/s41467-020-16123-w

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© The Author(s) 2020

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. 1 — a, b alexa568-phalloidin (Phalloidin) staining reveals actin accumulation in the well-organised muscle sheet around the gland epithelium. b Higher magnification reveals actin bridges between the muscle fibres. This muscle sheet defines a stromal compartment in the gland. c–f Viking-GFP (GFP tagged Collagen IV, Vkg-GFP in (c, d) or Perlecan-RFP (Perlecan in (e, f)) expression reveals the presence of a basement membrane, and Fasciclin III (c, d) or Coracle (e, f) staining reveals the apico-lateral domain of the epithelial cells. Transversal optical section (along the yellow line in (c) and (e)) confirms deposition of Viking-GFP and Perlecan-RFP at the basal pole of the epithelial compartment. g, h Transversal optical section of the gland at high magnification. Viking-GFP expression reveals the presence of the basement membrane, E-Cadherin (Ecad) staining reveals the apico-lateral domain of one epithelial cell and alexa633- phalloidin (Phalloidin) staining reveals actin accumulation in the muscle fibres. Arrows indicate where the basement membrane encloses the muscle fibres. DAPI (blue) reveals nuclei in (a) and (b)–(h), mostly from binucleated epithelial cells (see g–h). Representative images in (a–d, g, h) from three or more experiments and in (e–f) from two. Scale bars: 50 μm in (a–f) and 10 μm in (g, h).