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. 2023 Aug 30;621(7978):404–414. doi: 10.1038/s41586-023-06496-5

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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 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. 2 — a, FLT3^N399D or FLT3^N399G cells avoid 4G8-CAR-mediated killing. K562 cells expressing FLT3 variants were cultured with 4G8-CAR at different effector:target ratios (E:T). Left, FACS analysis of K562 cells expressing either FLT3 WT, N399D or N399G, after 48 h of co-culture. T cells and targets are identified by CD3 and FLT3, respectively. From left to right, the fraction of live target cells (absolute (abs.) counts of annexin V⁻7-AAD⁻ cells) relative to E:T = 0; T cell activation by CD69⁺ (%) and surface expression of FLT3 by BV10A4 staining on residual live targets, normalized (norm.) to E:T = 0. The E:T ratio is reported on the x axis. Data are mean ± s.d. n = 4. Statistical analysis was performed using two-way analysis of variance (ANOVA); P values of the comparisons between each condition are reported. b, KIT^H378R cells avoid Fab-79D-CAR killing. Left, plots of K562 cells expressing wild-type (WT) KIT, H378R or unmodified (KIT⁻) (K562 WT) after 48 h of co-culture with Fab-79D CAR T cells. T cells and targets are identified by CD3 and KIT, respectively. From left to right, the fraction of live target cells; T cell activation on the basis of CD69 staining; and surface expression of KIT by 104D2 on residual live targets, normalized to E:T = 0. The E:T ratio is reported on the x axis. Data are mean ± s.d. n = 4. Statistical analysis was performed using two-way ANOVA. c, CD123^S59 base-edited cells are resistant to CSL362 CAR T cells. Left, representative plots of CD123-reporter cells, either unmodified (K562 WT) or base-edited, or CD123⁻ K562 cells after 48 h of co-culture with CSL362-CAR. T cells and targets are identified by CD3, CD4 and CD8, and CD123, respectively. From left to right, the fraction of live target cells; T cell activation on the basis of CD69 staining; and surface expression of CD123 on the basis of staining with the 6H6 control antibodies on residual target cells. The E:T ratio is reported on the x axis. Data are mean ± s.d. n = 6. Statistical analysis was performed using two-way ANOVA.

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