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. 2019 Nov 13;10:5151. doi: 10.1038/s41467-019-13086-5

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© The Author(s) 2019

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. 3 — CR-31 translationally targets glutathione metabolism in PDA. a Heatmap of glutathione metabolism pathway proteins (KEGG database) showing fold-changes of polysome-associated mRNA. b Relative levels of NADPH normalized to viability based on CellTitre-Glo (CTG) assay (left) and ratio of NADP⁺ to NADPH (right) in organoids and patient-derived PDA lines upon treatment with vehicle (DMSO) or 100 nM CR-31 for 16 h. Data are mean +/–S.D. Student’s t-test, n = 4 (organoids), n = 5 (patient-derived lines). NS = not significant. c Relative levels of glutathione in patient-derived PDA lines upon treatment with vehicle or CR-31 for 24 h. Data are mean +/–S.D., Student’s t-test. n = 5, NS = not significant. *p < 0.05 compared to DMSO treatment. d Ratio of GSH to GSSG in patient-derived PDA lines upon treatment with vehicle or the indicated concentrations of CR-31 for 24 h. Data are mean +/–S.D., Student’s t-test, n = 5. *p < 0.05 compared to DMSO treatment. e ROS levels in KP organoids upon treatment with vehicle or 10 nM CR-31 for 16 h. Data are representative from three biological replicates. f NRF2 protein levels in KP organoids treated with CR-31 for 16 h. Actin, loading control. Representative image from 3 biological replicates. g mRNA expression of NRF2 target genes in 3 patient-derived PDA lines upon treatment with CR-31 for 5 or 24 h. Data are mean +/–S.D. Each data point plotted is average from every biological replicate. h Cell viability of KP and Nrf2-deficient KP (KPn) organoids upon a 72 h treatment with increasing concentrations of CR-31. Dotted lines indicate 95% confidence intervals, n = 5. i Cell viability of patient-derived PDA lines upon treatment with increasing concentrations of CR-31 in the presence or absence of 100 μM buthionine sulfoximine (BSO) for 72 h. Data are mean +/–S.D., n = 5. j Cell viability of primary murine PDA lines upon treatment with increasing concentrations of CR-31 in the presence or absence of 1 mM of the ROS scavenger N-acetylcysteine (NAC) for 72 h. Data are mean +/–S.D., n = 3. Source data are provided as a Source Data file