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. 2022 Mar 4;21:66. doi: 10.1186/s12943-022-01541-9

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© The Author(s) 2022, corrected publication 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/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 1 — Generation of CAR T cells targeting FLT3 and NKG2DLs. A A schematic representation of the manufactured FLT3scFv/NKG2D-CAR lentiviral construct. SP, signal peptide; scFv, single-chain fragment variable; Hinge, hinge chain. B FLT3scFv (EB10) and NKG2D expression assessed by flow cytometry of GFP + -transfected 293 T cells (B-1) and GFP + CAR T cells (B-2). The histograms show staining with recombinant FLT3 protein + anti-His APC antibody or anti-human NKG2D antibody (blue) compared to staining with isotype (red). C FLT3scFv (EB10) and NKG2D expression assessed by real-time PCR in transfected 293 T cells (C-1) and CAR T cells (C-2) compared with untransfected 293 T cells and UTD cells, respectively. D Significant CAR T cell transfection efficiency indicated by the percentage of GFP⁺ cells determined by flow cytometry. E CD4 and CD8 subtypes of CAR T cells assessed by flow cytometry. The data shown are derived from three independent experiments with T cells from different donors. UTD, un-transduced CD3 + T cells; GFP, green fluorescent protein; SSH, side scatter height; APC, allophycocyanin; PE, phycoerythrin. * p < 0.05, **** p < 0.0001