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. 2020 Nov 18;11:5883. doi: 10.1038/s41467-020-19659-z

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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. 5 — a Maximum intensity projection of fluorescence image from live 3D confocal imaging of Alexa-647 fibronectin NMBS (Magenta, 2 µm × 2 µm × 2 µm) applied to a developing Drosophila ovariole expressing moesin-GFP (Green) and Histone-2B-RFP (Red). b 3D render of a diagonal cut plane demonstrating the ability of the NMBS to conform to the 3D surface of the ovariole. c Strain analysis and quantification of region in a (white box) at two time points show regionally localized tensile (red) and compressive (blue) strain over time. d Magnified images (white box in c) of high tensile region (white arrows) corresponding to localized moesin-GFP fluorescence (green, yellow arrows). e Quantification of the average tensile strain (red, white arrows) within the magnified region shows strain peaks correlate with increased moesin-GFP fluorescence (green, yellow arrows) over a 4-h time course (mean S.D.; n = 5 segments over 1 experiment). f Maximum intensity projection of fluorescence image from live high-speed spinning-disk confocal imaging of Alexa-647 fibronectin NMBS (Magenta, 2 µm × 2 µm × 2 µm) applied to a contracting Drosophila ovariole expressing moesin-GFP (green) and Histone-2B-RFP (red). g Time-lapse imaging of region in f (white box) at 0.5 Hz of a 20 µm section of a contracting ovariole with the NMBS (magenta) and histone-2B-RFP (green). Regions of tension (t = 10 s, max of 1.55) and compression (t = 12 s, max of 0.63) are observed during contraction and extension respectively. As the nuclei translate upward during ovariole contraction (t = 10 s; red dots, gray commit tails) there is an increase in tension at their leading edge (max tension = 1.44). As the nuclei move down and the ovariole compresses (t = 12 s), a band of compression along the bottom edge of the ovariole is observed (max compression = 0.60). h Magnified image of strain analysis showing the 3D deformation during ovariole contractions and highlighting regions of compression and tension for quantification. i Quantification of the maximum tensile and compressive regions within the strain map, (ROI 1, magenta; ROI 2, cyan) of the contracting ovariole. Stress during contraction was calculated by using values for the elastic modulus of the germarium and stage 1 ovariole of 2 kPa (mean ± S.D.; ROI 1 n = 10 segments and ROI 2 n = 14 segments over 1 experiment).