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. 2022 Feb 22;126(12):1824–1833. doi: 10.1038/s41416-022-01742-5

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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. 5 — a Organoids derived from control (ECAS) and two independent GCLC-knockout clones (KO1 and KO2) were seeded in an NAC-depleted growth medium with or without reduced glutathione (GSH, 2 mM) as indicated (n = 3 per condition). Cells were treated with oxaliplatin for 72 h at the indicated concentrations (μM), and cell viability was then measured using Titer-Glo assay. The bar graphs show the percentage of surviving cell fractions. All values are plotted as % of untreated controls. b As in (a), with 5-FU or irinotecan, as indicated. (n = 3 per condition). c Control and GCLC-knockout organoids were exposed to oxaliplatin (5 h; 200 μM) in the presence or absence of reduced GSH (2 mM). After extensive washing of the cells, genomic DNA was isolated and the platinum (Pt) content was quantified by HPLC. The barplot shows the Pt content (n = 3) in ng per ug total genomic DNA. d Western blot analysis of pRPA, GCLC and actin protein expression levels in control ECAS and GCLC-knockout organoids following exposure to oxaliplatin (200 μM) at the indicated time points.