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. 2022 Aug 8;7(15):e150079. doi: 10.1172/jci.insight.150079

Figure 3. Prolonged stimulation through activating receptors weakens natural cytotoxicity.

Figure 3

(A) NK cells harvested from isotype-coated and exhaustion plates (day 7) were incubated with K-562 targets for 48 hours (E/T: 2:1). K-562 cells were labeled with CellTrace Far Red, and a fluorescent caspase-3/7 reagent was added to detect apoptotic cells. Killing was measured in real time using the IncuCyte live cell killing platform. (B) Percentage live targets was quantified for both groups of cells at multiple time points (n = 8). (C–E) NK cells were harvested from isotype-coated and exhaustion plates (day 7), and expression (MFI or % of parent) for granzyme B (C) (n = 9), perforin (D) (n = 7), and TRAIL (E) (n = 4) was measured via flow cytometry at the noted time points. Paired t tests were used for comparisons. *P ≤ 0.05; **P < 0.01; ***P < 0.001.