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. Author manuscript; available in PMC: 2022 Mar 24.
Published in final edited form as: Sci Immunol. 2021 Sep 24;6(63):eabe6968. doi: 10.1126/sciimmunol.abe6968

Fig. 5. NKG2C+CD8+ T cells can be generated in vitro by BCL11B deletion.

Fig. 5.

(A) FACS histograms showing BCL11B intracellular expression on CD8+ T cells electroporated with Cas/9 and control mRNA guides or BCL11B specific mRNA guides at the indicated time points. (B) FACS plots showing the frequency of CD8+CD56+ cells on the control and BCL11B KO cells after 2 or 4 weeks of expansion with CD3/CD2/CD28 beads or K562 mbIL21 HLA-E:VMAPRTLFL. Graph on the right shows cumulative analysis from 2 independent donors. (C) FACS plots showing the frequency of CD45RA+CCR7−CD8+ cells on the control and BCL11B KO cells after 2 or 4 weeks of expansion with CD3/CD2/CD28 beads or K562 mbIL21 HLA-E:VMAPRTLFL. Graph on the right shows cumulative analysis from 2 independent donors. (D) Cumulative analysis from 2 independent donors on PD-1+CD8+ T cells in control or BCL11B KO cells after 2 or 4 weeks of expansion with CD3/CD2/CD28 beads or K562 mbIL21 HLA-E:VMAPRTLFL. (E) FACS plots showing the frequency of NKG2A+ vs NKG2C+ and CD56+DAP12+CD8+ T cells on the control and BCL11B KO cells after 4 weeks of expansion with K562 mbIL21 HLA-E:VMAPRTLFL. (F) Cumulative analysis of 2 independent donors from the data shown in (E). (G) Representative flow cytometry plots showing degranulation (CD107a) and intracellular IFN-γ expression by K562 mbIL21 HLA-E:VMAPRTLFL-expanded pre-gated NKG2C+ or NKG2C−CD8+ T cells after 6 hours stimulation with the indicated target cells.