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. 2019 Jun 13;34(1):100–114. doi: 10.1038/s41375-019-0507-8

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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 — ND2158 exerts preferential cytotoxicity for CLL cells. a Viability was analyzed by flow cytometry after 48 h of incubation of cells in culture at ND2158 concentrations from 10 to 100 µM in MYD88-mutated IGHV-mutated (n = 6), MYD88-unmutated IGHV-mutated (n = 16) CLL samples, and in CD19⁺ B cells and CD3⁺ T cells from healthy donors (n = 10). Percentage of viable cells was measured by Annexin-V and normalized to untreated control. b Viability of ND2158-treated CLL cells was analyzed after TLR stimulation for 2- (n = 12) and 6 days (n = 6). c Left panel: Percentage of proliferating CD19⁺ CLL cells after TLR stimulation and ND2158 treatment for 6 days measured by CFSE dilution (n = 5 MYD88-mutated; n = 7 MYD88-unmutated). Right panel: Flow cytometry histogram of a representative MYD88-unmutated CLL case (#51) shows proliferating cells (gated on viable CD19⁺ cells) after 6 days of TLR and IL15 stimulation and ND2158 treatment. A decrease in CFSE signal is indicative for cells that have divided. Wilcoxon signed-rank test was used for statistical analysis. Horizontal bars represent population means. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. M mutated, UM unmutated