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. 2023 Feb 2;14:563. doi: 10.1038/s41467-023-36160-5

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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 Workflow for CD133⁺ CAR-T production and UNC0642 treatment. Created with BioRender.com. B Granzyme B expression levels in CAR-T cells after UNC0642 treatment (n = 3 donors). C Example normalized cell index for 2D target cell killing assay for untreated and UNC0642-treated CD133⁺ and mCH (CD133^-) CAR-T cells. CAR-T cells were added to target cells at a 1:4 effector: target ratio (n = 3 donors). D Area under the curve for normalized cell index as show in C for untreated and UNC0642-treated CD133⁺ and mCH CAR-T cells at 1:2, 1:4 and 1:16 effector:target ratios. Data are analyzed with one-way ANOVA with multiple comparisons (n = 3 donors). E 3D target cell killing assay for untreated and UNC0642-treated CD133⁺ and mCH CAR-T cells at day 2 after CAR-T addition. Data are normalized to no T cell target cell numbers. Data are analyzed with one-way ANOVA with multiple comparisons. N = 9 analyzed ROI regions from three biologically independent donors. F CAR-T cell migration in 3D target cell killing assay for untreated and UNC0642-treated CD133⁺ and mCH CAR-T cells at day 2. Data are analyzed with one-way ANOVA with multiple comparisons. N = 9 analyzed ROI regions from three biologically independent donors. All data are shown as mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Source data are provided as a Source Data file.