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. 2023 Aug 4;14:4677. doi: 10.1038/s41467-023-39571-6

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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. 7 — a Phospho-Kinase Array (n = 1 experiment) revealed that asparagine addition (2 mM, 0.5 h, 4 h) in glutamine-low (0.1 M, 12 h) medium activated p-SRC and validation by Western blot (n = 3 independent biological replicates). b SLC25A22 knockout suppressed p-SRC in DLD1 and CT26 cells (n = 2). c Glutamine (1 mM, 12 h) promoted p-SRC in an SLC25A22-dependent manner. Asparagine (2 mM, 12 h) restored p-SRC in SLC25A22 knockout cells (n = 3 independent biological replicates). d Asparagine increased thermal stability (5 min incubation) of recombinant SRC (n = 3 independent biological replicates). e BIAcore analysis of the binding of asparagine to recombinant SRC (n = 2). f Asparagine-induced recombinant SRC phosphorylation and kinase activity (n = 4). Each dot represents an independent sample. g Overlapping of in silico prediction of CXCL1 transcription factors (TFs), SRC-downstream TFs, and SLC25A22 knockout RNA-seq data revealed ETS2 as a putative transcription factor for CXCL1. h SLC25A22 knockout suppressed ETS2 and p-ETS2 expression, and i ETS2 nuclear localization in DLD1 and CT26 cells (n = 2). j Glutamine (1 mM, 12 h) increased ETS2 protein in a SLC25A22-dependent manner. Asparagine (2 mM, 12 h) rescued ETS2 in SLC25A22 knockout cells (n = 2). k ASNS knockdown suppressed ETS2 expression (n = 2). l ETS2 knockdown reduced CXCL1 mRNA and secretion (n = 3). Each dot represents an independent sample. m ETS2 overexpression rescued CXCL1 mRNA and secretion in SLC25A22 knockout cells (n = 3). Each dot represents an independent sample. n In silico identification of ETS2 binding sites on CXCL1 promoter and validation by ChIP-PCR. o Luciferase reporter assay confirmed that ETS2 activated transcription of CXCL1 promoter (n = 3). p SRC inhibitor Bosutinib (24 h) suppressed ETS2, p-ETS2 expression, and CXCL1 secretion in DLD1-sgControl cells, without any effect in SLC25A22 knockout cells (n = 3). Each dot represents an independent sample. Data are shown as mean ± SD (f, l, m, o, p). Two-tailed one-way ANOVA (f, l, p). Two-tailed Student’s t test (m, o). Source data are provided as a Source Data file.