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. 2019 Jul 12;9:10127. doi: 10.1038/s41598-019-46470-8

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© The Author(s) 2019

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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Cell death in the abdominal FB cells of 120QHTT^ex1 adults. (a) Representative bright-field images of abdominal FB from day10 and day15 old 25QHTT^ex1 and 120QHTT^ex1 females show dramatically degenerated FB in 120QHTT^ex1 females. Scale bar, 100 µm. (b) Acridine orange (AO) staining was performed on live abdominal FB tissue of both 25QHTT^ex1 and 120QHTT^ex1 females throughout the indicated ages (day0-day15). AO positive regions, an indication of cell death, are visible as bright spots (green) of higher intensity throughout the FB in 120QHTT^ex1 flies. Abdominal FB of 120QHTT^ex1 flies shows fewer AO positive regions at day5 while a dramatic increase in the proportion of AO positive areas is observed afterwards till day10 of age. At day15, AO signal is slightly reduced than day10 with severe FB degeneration. (n = 6–8/genotype/age) Scale bar, 10 µm. (c) Histogram representing the mean fluorescence intensity (measured in arbitrary units) of AO in abdominal FB of 25QHTT^ex1 and 120QHTT^ex1 females. (d) Visualisation of cell death in abdominal FB from 5- and 15- days old 25QHTT^ex1 and 120QHTT^ex1 females by TUNEL labelling. TUNEL-positive cells are stained green and nuclei are labelled with DAPI. TUNEL-positive cells are detected in large amounts in the FB of day15 old 120QHTT^ex1 adults. 25QHTT^ex1, however, do not show TUNEL-positive cells. (n = 8–10/genotype/age) Scale bar, 10 µm. (e) Histogram representing the mean intensity of TUNEL signal (green; measured in arbitrary units) in abdominal FB of 25QHTT^ex1 and 120QHTT^ex1 females. Values represent mean ± SEM. *** P < 0.001; **P < 0.01.