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. 2023 Jun 8;14:3375. doi: 10.1038/s41467-023-39097-x

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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. 5 — a, b OSCAR T cell cytotoxic activity measured as TNFα production and expression on CD8 (top) and CD4 (bottom) T cells upon co-culture with different normal tissues. TNFα expression is detected by flow cytometry intracellular staining at 6 h co-incubation at an E:T ratio of 1:2. a OSCAR T cell cytotoxicity against human MSCs and MSC-derived osteoblasts at different stages of differentiation. Data are mean ± s.d. pooled from three independent experiments (n = 3). OSCAR T cells are from three healthy donors. Human MSC are from two donors. Statistical comparisons were performed with two-way ANOVA with Tukey’s multiple comparison test (ns not significant p > 0.05, *p < 0.05, ****p < 0.0001). b OSCAR-1 and OSCAR-3 T cell reactivity against healthy tissue cells representing lung (MRC-5, Hulec-5a, HPAEpiC), liver (HH), and kidney (HREpC). Data are mean ± s.d. pooled from three independent experiments (n = 3). OSCAR T cells are from three healthy donors. Statistical comparisons were performed with two-way ANOVA with Tukey’s multiple comparison test (ns not significant p > 0.05, ****p < 0.0001). c ALPL and TP-3 protein expression in different tumor cell lines from osteosarcoma (OHS, OSA, SaOS-2, LM7, U2OS), other tumors (MCF-7, MDA-MB-231, SK-OV-3, HeLa, HCT116, Caco-2, LS174T, U87, U251, U251mg, EBV-LCL, Rec-1, U-2932, BL-41, K562, THP-1, PANC-1, HEK, PC-3) and cells from different healthy tissues (BJ, HUVEC), distinct cell types from PBMCs (n = 3), and BM (n = 3, except HSCs n = 1). Results were normalized by the ratio between the Mean Fluorescence Intensity (MFI) obtained after staining with ALPL (green) or TP-3 (blue) and that of the appropriate control, before log2 transformation. U2OS is empty circle. d OSCAR-1 and OSCAR-3 T cell reactivity versus hematopoietic stem and progenitor cells (HST) from healthy bone marrow by colony-forming unit (CFU) assay. OSCAR T cells and HST were co-cultured at a 10:1 E:T ratio. Data represents the number of counted colonies; white, red, and total (mixed) colonies. Data are shown as mean ± s.d. pooled from two independent experiments with two donors (n = 2).