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. 2023 Dec 18;14:8405. doi: 10.1038/s41467-023-44098-x

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© The Author(s) 2023

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 Schematic of the key roles of the pentose phosphate pathway. Analysis and quantification of Thioltracker (GSH, b–d) and secretory activity (e) in G6PD-RNAi and PDG-RNAi tubules (driven by CapaR-Gal4). f, g WGA and DAPI staining in G6PD-RNAi tubules at 7 days and 1 day (h, i) post-eclosure. j, k Quantification of nuclear area (DAPI) and cell area. l–n H3K9me3 and DAPI, Lysotracker o–q and SA β-Gal (r–t) staining and quantification in 7-day G6PD-RNAi tubules. Data represented as box and whisker plots (lower and upper hinges correspond to the first and third quartiles, median line within the box, whiskers extend from the hinge to the largest/smallest value, at most 1.5* interquartile range of the hinge) with all data from MpT main segment sections (d, q, t), nuclei/cells from main segments (j, k, n) or secretion of individual kidneys (e) shown as overlaid points. NS Not Significant, **P < 0.01, ***P < 0.001 (unpaired two-tailed t-tests or ANOVA followed by Tukey multiple comparisons test where appropriate). p values: (d) p < 0.0001, (e) control vs G6PD p = 0.00507, control vs PGD p < 0.0001, (j) 1 day control vs 7 day G6PD p < 0.0001, 1 day G6PD vs 7 day G6PD p < 0.0001, 7 day control vs 7 day G6PD p < 0.0001, (k) 1 day control vs 7 day control p = 0.00514, 1 day control vs 7 day G6PD p < 0.0001, 7 day G6PD vs 1 day G6PD p < 0.0001, 7 day G6PD vs 7 day control p < 0.0001, (n) p < 0.0001, (q) p = 0.000567, (t) p = 0.000574. p values where p > 0.05 labelled as NS. For analysis of fluorescent reporters/dyes, two images of different sections of the MpT main segment per fly were imaged. All images representative of >5 tubules. All images are maximum z projections. d n = 10 tubules per condition, (e) n = 29 control, 15 G6PD and 10 PGD tubules, (j) 73 nuclei (from 6 control 1 day tubules), 66 nuclei (from 4 G6PD 1 day tubules), 68 nuclei (from 5 control 7 day tubules) and 97 nuclei (from 9 G6PD 7 day tubules), (k) 70 cells (from 6 control 1 day tubules), 61 cells (from 4 G6PD 1 day tubules), 50 cells (from 5 control 7 day tubules) and 45 cells (from 9 G6PD 7 day tubules), (n) n = 56/50 nuclei (from 4 tubules) per condition, (q) n = 6 tubules per condition, (t) n = 6 tubules per condition. Source data are provided as a Source Data file.