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. 2020 May 8;11:2316. doi: 10.1038/s41467-020-15847-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. 4 — a Targeted metabolomics of livers of animals fed a Western-style diet. For each metabolite and each EDC-exposed animal, we computed its z-score (number of standard deviations) compared to the vehicle-treated animals. N = 5 biologically independent animals per treatment. The results were visualized with a CBioPortal Oncoprint-type exploration tool, and metabolites increased by at least 1.5 standard deviations in at least 3 animals and down in none or decreased by at least 1.5 standard deviations in at least 3 animals and up in none are shown. Genes that encode enzymes associated with altered metabolites are depicted on the left in italics (Ahcy, Mthfd1, Chdh, and Shmt1). S-Adenosylhomocysteine = SAH, homocysteine = HCY, S-ribosyl homocysteine = SRH, tetrahydrofolate = THF, phosphoenolpyruvic acid = PEP, fructose-bisphosphate/glucose-bisphosphate = FBP/GBP, glycine = Gly, leucine = Leu. b Metabolism set enrichment analysis (MetSEA) using genes identified by RNA-seq (shown in Fig. 3a) and metabolites identified in (a) to elucidate metabolic pathways disrupted in EDC-reprogrammed liver. MetSEA was performed using integrated recurrently up- or downregulated metabolites from livers of EDC-exposed animals with transcriptional signatures using the hybrid metabolite/transcriptomics pathway compendium WikiPathways. ORA using a hypergeometric distribution against WikiPathways identified 14 significantly enriched pathways at q < 0.2. c RT-qPCR validation of differentially expressed genes in key metabolic pathways identified by RNA-seq and MetSEA. Data from an independent validation set of vehicle-exposed (VEH; black dots) and EDC-exposed (EDC; red squares) animals fed a Western-style diet are shown. N = 9 biologically independent animals for VEH and N = 10 biologically independent animals for EDC. The p values generated by t test are indicated. *p ≤ 0.05; **p < 0.01; ***p < 0.001. Source data for c are provided as a Source data file.