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. 2021 Jun 8;22(8):e52032. doi: 10.15252/embr.202052032

Figure 1. ArCK cells are recognized by natural killer (NK) cells in vitro .

Figure 1

  1. Schematic representation for the generation of ArCK cells. Time 0 is defined by the estimated onset of Mps1 inhibitor‐induced chromosome mis‐segregation.
  2. Representative images of euploid control or ArCK cells interacting with NK cells. The NK cell‐mediated killing was measured at a 2.5:1 (NK cells: target cells) ratio and was recorded by live cell imaging for 36 h at a 30‐min interval. TO‐PRO‐3 (1 μM) was added to the medium at the same time of NK cell addition to measure cell membrane integrity. Phase contrast (top) and TO‐PRO‐3 signal (bottom) from the same field were presented. Arrowheads indicate ArCK cell death. All images were acquired at the same exposure time and light intensity. Scale bar 20 μM.
  3. Measurement of NK cell‐mediated killing of ArCK and euploid control cells (Ctrl) at a 2.5:1 (NK cells: target cells) ratio. 50 randomly chosen target cells were followed for 36 h by live cell imaging per condition per replicate. The cumulative cell death was calculated. n = 3 biological replicates; mean ± SEM. The statistical significance was determined using nonparametric Kolmogorov–Smirnov test (KS test) as described in the method section; P < 0.0001.
  4. Measurement of euploid control (Ctrl) and ArCK cell proliferation without NK cells. Live cell imaging of target cells without NK cells was performed using the same condition as described in the method section. For each condition, 50 cells were randomly chosen at the beginning of the movie as the initial population (indicated by the dashline, n initial = 50). The cumulative cell number was recorded. Dot plot of individual data points and mean was presented; n = 2 biological replicates.
  5. NK cell‐mediated cytotoxicity across various NK cell‐to‐target cell ratios. Either euploid control (Ctrl) or ArCK cells were co‐cultured with NK cells at the indicated NK cell: target cell ratios. The cumulative killing of target cells was measured. n = 3 biological replicates; mean ± SEM. P < 0.0001 for all four NK cell: target cell ratios, KS test.