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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. 2 — a Threonine is metabolized by threonine dehydrogenase (TDH) to 2-amino-3-ketobutylate, which then either spontaneously decarboxylates into aminoacetone or is metabolized by the GCAT enzyme using CoA as a co-substrate to form glycine and acetyl-CoA. b Threonine abundance decreased with age (***p < 0.001 versus the first time point, n = 3 worm pellets). c Glycine and homoserine decreased in DR or eat-2 nematodes (***p < 0.001 versus N2 group, n = 3 of 5-day-old-worm pellets). d Venn analysis of DR and eat-2 mutant using publicly available datasets. Intersection of differentially expressed genes (DEGs) of DR (p < 0.05 in edgeR, Deseq) with eat-2 mutant DEGs compared to the wild-type N2 (p < 0.05). e–g Among the common DEGs, the expression of enzymes that were related with threonine metabolism were determined as (e) R102.4 (**p < 0.01 and ***p < 0.001 versus the N2 group, n = 3 of 5-day-old-worm pellets), (f) TDH/F08F3.4 (***p < 0.001 versus the N2 group, n = 3 of 5-day-old-worm pellets), and (g) K01C8.1 (p > 0.05, n = 3 of 5-day-old-worm pellets). The expression of these enzymes was measured with age as (h) R102.4 (**p < 0.01 versus the first time point, n = 3 worm pellets), (i) TDH/F08F3.4 (**p < 0.01 and ***p < 0.001 versus the first time point, n = 3 worm pellets), and (j) K01C8.1 (p > 0.05). k Lifespan analysis of tdh/F08F3.4 RNAi (mean ± s.d, p < 0.001, log-rank test, n = 88–96 worms with 3 independent assays) versus control RNAi in N2 worms. l–r Effects of tdh/F08F3.4 RNAi versus control RNAi for (l) threonine contents (***p < 0.001, n = 3 worm pellets), (m) average speed (***p < 0.001, n = 10–15 worms with 3 independent assays), (n) body length (p = 0.785, n = 6), (o) triglyceride (TG) content (***p < 0.001, n = 3 worm pellets), (p) superoxide dismutase (SOD) activity (**p = 0.007, n = 3 worm pellets), (q) catalase activity (**p = 0.002, n = 3 worm pellets), and r oxidative stress resistance (***p < 0.001, n = 20 worms × 9 measurements each). b–c, e–j and m–r P values were determined by two-sided t-test. FDR < 0.05. Error bars represent the mean ± s.d. Source data are provided as a Source Data file.