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. 2021 Oct 26;12:6176. doi: 10.1038/s41467-021-26395-5

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© The Author(s) 2021

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. 1 — A Growth curve of A459, HCT116 and MDA-MB-231 grown in DMEM and Plasmax with or without serine/glycine (SG-). Data are represented as mean ± SD of triplicate wells. B Intracellular levels of serine measured by LC–MS after incubation with 400 µM or 140 µM of 2,3,3-D₃-Serine in DMEM and Plasmax, respectively. Data are represented as mean ± SD of triplicate wells. C Serine is converted by the SHMT enzymes into glycine which will contribute to a number of important processes such as redox defence, nucleotide production and provision of one-carbon units to the folate cycle and methylation reactions. D Extracellular formate measured after 72 h in DMEM (grey) and Plasmax (blue) by NMR. Data are represented as mean ± SD of duplicate wells. E Intracellular levels of glycine measured by LC–MS after 4 h incubation with 400 µM or 140 µM of 2,3,3-D₃-Serine in DMEM or Plasmax, respectively. Data are represented as mean ± SD of triplicate wells. F Intracellular levels of GSH measured by LC–MS after 4 h incubation with 400 µM or 140 µM of 2,3,3-D₃-Serine in DMEM or Plasmax, respectively. Data are represented as mean ± SD of triplicate wells. G Intracellular levels of GSH measured by LC–MS after 4 h incubation with 400 µM or 330 µM of 2,2-D₂-Glycine in DMEM or Plasmax, respectively. Data are represented as mean ± SD of triplicate wells. H Intracellular levels of dTTP measured by LC–MS after 4 h incubation with 400 µM or 140 µM of 2,3,3-D₃-Serine in DMEM or Plasmax, respectively. Data are represented as mean ± SD of triplicate wells. I Intracellular levels of AMP and ATP measured by LC–MS after 4 h incubation with 400 µM or 140 µM of 2,3,3-D₃-serine in DMEM or Plasmax, respectively. Data are represented as mean ± SD of triplicate wells. J Intracellular levels of ATP measured by LC–MS after 4 h incubation with 400 µM or 330 µM of 1-¹³C-glycine in DMEM or Plasmax, respectively. Data are represented as mean ± SD of triplicate wells. All graphs are representative of 3 independent experiments. Source data are provided as a Source Data file.