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. 2022 Nov 2;13:6554. doi: 10.1038/s41467-022-34265-x

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© The Author(s) 2022

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. 4 — a–d Lifespan assay of tdh/F08F3.4 RNAi (red) versus control RNAi (black) with transcripts of DAF-16 or HSF-1 among the commonly regulated genes of tdh/F08F3.4 RNAi, DR, and eat-2 (by Venn analysis, Supplementary Data 3), (a) gst-19/F37B1.8 RNAi, (b) T12D8.5 RNAi, (c) cnc-2/R09B5.3 RNAi, and d ftn-1/C54F6.14 RNAi worms. e Lifespan assay of ftn-1(ok3625) with tdh/F08F3.4 RNAi (red-dashed) versus control RNAi (black-dashed) compared with survival curves of N2 (bold-line). f Effects of threonine (100, 200, and 400 μM) versus the vehicle (black) on lifespan of the ftn-1(ok3625) mutant. P value and lifespan assay data summarized in Supplementary Table 1. g Expression of ftn-1 under tdh/F08F3.4 RNAi was determined at intervals across lifespan (*p < 0.05 and ***p < 0.001 versus control RNAi, n = 3 worm pellets). h Expression levels of ftn-1 under ftn-1/C54F6.14 RNAi (#p < 0.001 compared to control RNAi, ***p < 0.001 versus ftn-1 RNAi, n = 3 worm pellets). i Quantification of Fe²⁺/Fe³⁺ iron contents of nematode with ftn-1/C54F6.14 RNAi at intervals across lifespan (#p < 0.001 compared to control RNAi, **p < 0.01 and ***p < 0.001 versus the ftn-1 RNAi, n = 3 worm pellets). j Relative Amplex Red fluorescence in supernatant of worms (#p < 0.001 compared to control RNAi, *p < 0.05 and **p < 0.01 versus ftn-1 RNAi, n = 3 worm pellets). k Total glutathione (GSH) level was normalized to the GSH level in worms exposed to tdh/F08F3.4 RNAi or threonine (#p < 0.001 compared to control RNAi, *p < 0.05 and **p < 0.01 versus ftn-1 RNAi, n = 3 worm pellets). l Levels of the malondialdehyde (MDA), were measured and normalized against the mean of control RNAi-treated worms for independent samples (#p < 0.001 compared to control RNAi, **p < 0.01 and ***p < 0.001 versus the ftn-1 RNAi, n = 3 worm pellets). m–o Effects of tdh/F08F3.4 RNAi or threonine with ferroptosis inducer erastin or diethyl maleate (DEM) to induce GSH depletion as relative levels of m ROS, (n) GSH, and o MDA with ferroptosis inhibitor, ferrostatin-1 (***p < 0.001 versus control RNAi, n = 3 worm pellets). Overall differences between conditions were analysed by two-way ANOVA. Differences in individual values or between two groups were determined using two-tailed t-tests (95% confidence interval). Error bars represent the mean ± s.d. Source data are provided as a Source Data file.