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. 2022 Jul 9;13:3984. doi: 10.1038/s41467-022-31762-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. 7 — a Glucose uptake analysis in control and Activin A (AC)-treated for 2 h EBs. The values are normalized to the protein concentration (n = 4 biological independent samples). b Glucose uptake analysis in Activin A (AC)-treated for 2 h or untreated WT and Cripto KD/Cryptic KD EBs. The values are normalized to the protein concentration (n = 3 biological independent samples). c Glucose uptake analysis in Activin A (AC)-treated for 2 h or untreated WT and Smad2 KO/Smad3 KD EBs. The values are normalized to the protein concentration (n = 3 biological independent samples). d Glucose uptake analysis in Activin A (AC- treated for 2 h or untreated WT and Trim33 KO EBs). The values are normalized to the protein concentration (n = 3 biological independent samples). e Glucose uptake analysis of Activin A (AC)-treated for 2 h or untreated WT and Nemep KO EBs at day 3. The values are normalized to the protein concentration (n = 3 biological independent samples). f Glucose uptake analysis of NEMEP-FLAG overexpressing WT and Nemep KO EBs at day 3. The values are normalized to the protein concentration (n = 3 biological independent samples). g Working model for the role of NEMEP during mesendoderm differentiation (Created with BioRender.com). A highly conserved putative lncRNA Gm11549 is specifically induced by Nodal signaling during mesendoderm differentiation and it is essential for mesendoderm differentiation in mESCs. There is a “ORF” hidden in Gm11549 that codes a transmembrane micropeptide, we named NEMEP. NEMEP functions by interacting with GLUT1 and GLUT3, and augment the glucose uptake capacity of glucose transporter proteins to meet the energy needs during mesendoderm differentiation. a–f: Data are the mean ± S.E.M. P values were determined by unpaired two-tailed t-tests (a), one-way ANOVA (e, f) with Dunnett’s corrections, two-way ANOVA (b–d) with Sidak’s corrections, and data are representative of three independent experiments with similar results. Source data are provided as a Source Data file.