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. 2022 Sep 27;72(4):917–928. doi: 10.1007/s00262-022-03290-6

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© The Author(s) 2022

Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — In vitro cytotoxicity of LCAR-M23 CAR T cells. a MSLN expression in six kind of cell lines was determined by flow cytometry. MSLN is expressed in OVCAR-3 cells, Caov-3 cells, and SK-OV-3 cells; MSLN is not expressed in A-431 cells, K-562 cells, and HEK-293 cells. b In vitro cytotoxicity of LCAR-M23 CAR T cells againstOVCAR-3 cells, Caov-3 cells, SK-OV-3 cells, A-431 cells, K-562 cells and HEK-293 cells. c Flow cytometry analysis of MSLN expression in ascitic cells of ovarian cancer patients. d In vitro cytotoxicity of LCAR-M23 CAR T cells against autologous ascitic tumour cells was determined by flow cytometry. e MSLN expression in engineered HEK-293 cell lines was determined by flow cytometry. f Cytotoxicity of LCAR-M23 CAR T cells against HEK-293 cells expressing gradient levels of MSLN. g IFNγ and TNFα release of LCAR-M23 CAR T cells in co-culture with HEK-293 cells expressing gradient levels of MSLN by using cytokine kits. The level of IFN γ and TNF α release of high MSLN clone LCAR-FM23 was significantly increased compared with only HEK-293 group, medium MSLN clone group and low MSLN clone group. Data shown in a–g are at least 3 experiments (mean ± SEM). *p < 0.05 ** p < 0.01, *** p < 0.001