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. 2018 Nov 5;9:4620. doi: 10.1038/s41467-018-06497-3

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© The Author(s) 2018

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 — Quantitative analysis of cell shape changes during fold formation. a Schemes representing top views (above) and cross-sectional views (below) of wing imaginal discs before and after folding. The type of fold is indicated. b–e Top view (b, d) and cross-sectional (c, e) images of a time-lapse movie of a cultured 72 h AEL wing imaginal disc expressing Indy-GFP, showing formation of hinge-hinge (H/H) and hinge-pouch (H/P) folds. Time relative to first appearance of apical indentation (AAI) (i.e. the first time when the apical surface of fold cells is below the apical plane of neighboring cells) of H/H fold is shown. In this and the following figures, top views are shown with dorsal to the left and posterior up; in cross sections, the apical surface of columnar cells is to the top, unless otherwise indicated. Dotted lines in top views indicate position of the corresponding cross sections. Scale bars are 10 μm. f, g Top view (f) and cross-sectional (g) images of the boxed areas of the time-lapse movie shown in b and d at indicated time points. Scale bars are 10 μm. h, i Schemes showing simplified cell shapes before and during folding and the set of geometric parameters used. d_a and d_b denote the apical and basal deformations, l_a and l_b denote the apical and basal cross-sectional lengths of cells located at the center of the fold, and h_tissue denotes the apico-basal height of cells neighboring the fold. j–m Cross-sectional images of a time-lapse movie of a cultured wing imaginal disc expressing Indy-GFP (gray) in all cells and RFP (turquoise) in clones of cells of H/H fold (j, k) or of H/P fold (l, m). Red dots mark apical and basal vertices of RFP-labeled cells. Scale bars are 10 μm. n–q Changes of the geometric parameters indicated in i during H/H (n, p) and H/P (o, q) fold formation as a function of time relative to AAI. All geometrical quantities are normalized by the cell height h_tissue of the surrounding tissue. Mean and s.e.m. are shown. n = 17 cross sections of 7 wing imaginal discs for n and p and n = 12 cross sections of 6 wing imaginal discs for o and q