Figure EV4. Mitochondrial morphology and functions of ITNKs.

1. Confocal microscopy images showing purified CD8⁺ T cells and CD8⁺NKp30⁺ ITNKs in which the mitochondria (MitoTracker; green) and nuclei (Hoechst; blue) are stained. Each dot represents the mean relative length of the 24 mitochondria per replicate. Scale bars: 5 μm.
2. Confocal microscopy images showing purified CD4⁺ T cells and CD4⁺NKp30⁺ ITNKs in which the mitochondria (MitoTracker; green) and nuclei (Hoechst; blue) are stained. Each dot represents the mean relative length of the 24 mitochondria per replicate. Scale bars: 5 μm.
3. ITNKs that were derived from T cells transduced with sgBCL11B, sgMTA2, sgMBD2, or sgCHD4 or sgCtrl‐transduced T cells were cultured for 10 days for observation of mitochondrial morphology. Confocal microscopy images are shown; mitochondria (MitoTracker; green); nuclei (Hoechst; blue), and NKp30 (purple) are stained. Scale bars: 5 μm.
4. CD4⁺NKp30⁺ and CD8⁺NKp30⁺ ITNKs derived from sgBCL11B‐transduced T cells were purified and assessed the mitochondrial functions by FACS. CD8⁺ T cells and CD8⁺NKp30⁺ ITNKs (left) and CD4⁺ T cells and CD4⁺NKp30⁺ ITNKs (right) were stained with MitoTracker green and analyzed by flow cytometry.
5. CD8⁺ T cells and CD8⁺NKp30⁺ ITNKs (left) and CD4⁺ T cells and CD4⁺NKp30⁺ ITNKs (right) were stained with TMRM and analyzed by flow cytometry.
6. CD8⁺ T cells and CD8⁺NKp30⁺ ITNKs (left) and CD4⁺ T cells and CD4⁺NKp30⁺ ITNKs (right) were stained with MitoSOX and analyzed by flow cytometry.

Data information: In (A, B, and D–F), data are presented as mean ± SD, N = 3 (A and B) or 6 (D–F) individual healthy donors. *P ≤ 0.05, **P ≤ 0.01, and ***P ≤ 0.001, two‐tailed paired Student's t‐test.

Source data are available online for this figure.