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. 2023 Sep 22;42(21):e113448. doi: 10.15252/embj.2023113448

Figure 1. Deletion of subunits of the NuRD complex reprograms human T cells into ITNKs.

Figure 1

  1. Immunoprecipitation of the nuclear extract of peripheral blood mononuclear cells (PBMC)‐derived T cells with anti‐BCL11B antibodies and western blot analysis with antibodies against the NuRD components MBD2, MTA2, and CHD4.
  2. Human T cells from PBMC were electroporated with sgCtrl, sgBCL11B, sgMBD2, sgMTA2, or sgCHD4 and Cas9 protein after activation with CD3/CD28 antibodies for 36 h. Those T cells were cultured in T cell culture medium containing rh‐IL2 (300 U/ml) for 10 days. Representative flow cytometric detection of NKp30 and NKp46 expression in PBMC‐derived T cells transduced with sgCtrl, sgBCL11B, sgMBD2, sgMTA2, or sgCHD4. The data shown is representative of four individual healthy donors.
  3. Graph summarizing the percentages of NKp30+ in CD4 T cells and the percentages of NKp46+ cells in CD8 T cells that were transduced with sgCtrl, sgBCL11B, sgMBD2, sgMTA2, or sgCHD4.
  4. After culture for 10 days, T cells in (B) were incubated with the K562‐GL target cells at various E:T ratios for 24 h. Killing assays showing the percent cytotoxicity of T cells transduced with sgCtrl, sgBCL11B, sgMBD2, sgMTA2, or sgCHD4 against K562‐GL cells. The data represents killing percentage of cells from a donor.

Data information: In (C and D), data are presented as mean ± SD (N = 4 individual healthy donors). **P ≤ 0.01, ***P ≤ 0.001 and ****P ≤ 0.001, one‐way ANOVA (C) or two‐way ANOVA (D) with Tukey's multiple comparisons test.

Source data are available online for this figure.