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. 2015 Dec 9;7(2):2070–2079. doi: 10.18632/oncotarget.6529

Figure 3. ML-2 cells reduces, in vitro, the frequency of short-term NK effector cells with anti-leukemia activity.

Figure 3

IL-2 post-activated STNK and LTNK cells were subjected to a 2 consecutive rounds of incubation with ML-2 cells at a ratio of 4 STNK or LTNK cells to 1 ML-2 cell at 37°C. Following a 5-hour and an 18-hour incubation of STNK and LTNK cells with ML-2 cells, cell aliquots were harvested and stained with FITC-annexin-V, PI, PE-anti-CD16, and APC-anti-CD56 and analyzed by flow cytometry for CD16 down-regulation panel A., NK cell apoptosis panel B., NK cell depletion panel C. and ML-2 cell damage panel D. Also, following the first incubation with ML-2 cells, STNK and LTNK cells were magnetically sorted from cell preparations and reincubated overnight with ML-2 cells at 37°C. Then, cells were stained with FITC-annexin-V and PI and analyzed by flow cytometry. ML-2 cells were identified by posting an electronic gate on cells with the highest forward and side scatters panel E.