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. 2023 Jan 4;6:8. doi: 10.1038/s42003-022-04404-7

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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. 3 — a Schematic of the Sprecher pathway of docosahexaenoic acid (C22:6n3) synthesis from alpha-linolenic acid (C18:3n3) through a series of elongation and desaturation steps in the endoplasmic reticulum (ER) that is finished within peroxisomes via their beta-oxidation machinery. b Gene expression analysis of the ER-localized Fatty acid desaturase 1 (Fads1) and 2 (Fads2) and Elongation of very-long-chain fatty acids-like 2 (Elovl2) and 5 (Elovl5) involved in the Sprecher pathway in the livers of three 2.5-month-old female WT and Tmem135^{FUN025/FUN025} (FUN025) mice. c Gene expression analysis of ATP binding cassette subfamily D member 2 (Abcd2) in the livers of three 2.5-month-old female WT and Tmem135^{FUN025/FUN025} mice. Ribosomal protein lateral stalk subunit P0 (Rlplp0) served as the housekeeping gene in these studies. d Western blot analysis of peroxisomal beta-oxidation enzymes including acyl-CoA oxidase 1 (ACOX1), D-bifunctional protein (DBP), acetyl-Coenzyme A acyltransferase 1 (ACAA1), and sterol carrier protein x (SCPx) using liver lysates from 2.5-month-old WT, Tmem135 TG (TG), and Tmem135^{FUN025/FUN025} mice. ACTB served as the loading control for these experiments. 4 WT (2 males/2 females), 4 Tmem135 TG (2 males/2 females), and 5 Tmem135^{FUN025/FUN025} (3 males/2 females) were used in these experiments. Asterisks (** and ****) indicates post hoc Tukey test for a P < 0.01 and P < 0.0001 significance following a significant difference detected by one-way ANOVA. Dots represent individual data points. The number in parentheses represents the N of independent mouse samples per genotype used in the experiment. The protein size next to the immunoblot images denotes the size of the immunoband measured for this analysis. These experiments were repeated twice to ensure reproducibility. Data are presented as mean ± SD.