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. 2023 Feb 20;14(2):144. doi: 10.1038/s41419-023-05677-4

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© The Author(s) 2023, corrected publication 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. 6 — A Heatmap of differentially accumulated metabolites between shNC and shSHMT2 ACHN cells based on the LC-MS metabolite profile. A represented shNC, B represented shSHMT2. B ROS labeled by DCF-DA were detected by flow cytometry after SHMT2 knockdown in ACHN cells. All values were normalized to siNC control values. C Cellular GSH/GSSG ratios were measured using a GSH/GSSG-Glo Assay kit. All values were normalized to siNC control values. D Representative confocal microscopy images of ACHN cells stained with Mito tracker (red) and DAPI nuclear staining (blue). Red fluorescence indicated the active mitochondria. E TEM analysis demonstrated the ultrastructure changes of mitochondria after SHMT2 knockdown in ACHN cells. Red arrows indicated mitochondria. The scale bars in the original image and the enlarged image represents 5 μm and 1 μm, respectively. F Quantification of the monomeric JC-1 levels, indicating a decrease in MMP after treatment with siSHMT2. G OCR measurements in ACHN cells after SHMT2 knockdown. H Basal respiration, estimated ATP production, and maximal respiration based on OCR results. I ECAR measurements in ACHN cells after SHMT2 knockdown. J Glycolysis, glycolytic capacity, and glycolytic reserve based on ECAR results. K Glucose uptake was measured using Promega glucose uptake-Glo assay kit.