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. 2016 Dec 19;215(6):875–889. doi: 10.1083/jcb.201604136

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© 2016 Hsu et al.

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Figure 5. — Nondirected degranulation outside of the IS increases bystander killing of the neighboring cells. Live-cell confocal microscopy of YTS-CD16 cells incubated with S2 cells as innocent bystanders and S2-IgG (A) or S2-IC1-IgG (B) cells as activating targets. NK cells were mixed with the target cells immediately before the imaging process and imaged every 5 min for 2 h. Yellow, IgG-labeled S2 or S2-IC1 cells; green, bystander S2 cells; red, LysoTracker red (lytic granules); blue, SYTOX blue viability dye. Arrowheads indicate uptake of SYTOX blue viability dye by the target/bystander cells. Quantitative analyses of nonspecific killing of S2 cells over total lysis are shown as a feature of collateral damage to the bystander S2 cells by YTS-CD16 (C) and eNK (D) cells. Each point indicates one independent experiment; n > 400 cells/group. Error bars show ± SD. **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.