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. 2020 Dec 14;11:6387. doi: 10.1038/s41467-020-19974-5

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© The Author(s) 2020

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 Chemical structures of IRE1 kinase (K) inhibitors. b IRE1 RNase (R) activity in the presence of compounds in panel a. RNA cleavage was measured by a kinetic fluorescence assay using an XBP1-like RNA hairpin substrate and IRE1 LKR construct (Q470-L977). Background from the no-protein control was subtracted from signal before calculating fold change. Source data are provided as a Source Data file. Data are presented as the mean for measurements from two independent experiments (n = 2). c RNA levels for IRE1 RNase product XBP1s measured by bDNA assay of lysates from KMS-11 cells treated with compounds for 4 h. Background from the no-RNA control was subtracted from signal before calculating fold change. Source data are provided as a Source Data file. Data are presented as the mean for measurements from two independent experiments (n = 2). d, e Analytical Ultracentrifugation sedimentation velocity (SV-AUC) experiments for (d), IRE1 LKR (Q470-L977) unphosphorylated and autophosphorylated and (e), IRE1 LKR unphosphorylated in the presence of slight excess G-9807 and G-1749. Calculated sedimentation coefficients for IRE1 monomer (3.1 svedbergs) and dimer (4.6 svedbergs) are highlighted by dotted lines. Source data are provided as a Source Data file.