Effects of birinapant on NK cells in primary samples from patients with CML. (A) Lineplot of NK-cell cytotoxicity at different E:T ratios in 5 CP and 6 BP patient samples. E:T ratio of 0 represents a condition containing no NK cells, only target CD34⁺ CML cells. CD34⁺ counts have been normalized to this no-NK control. (B) Box plot showing drug sensitivity profiling of primary CML CD34⁺ cells’ sensitivity to NK cells at different E:T ratios with and without drug treatment ex vivo. Dots represent normalized CML CD34⁺ cells of all CP and BP patients after 24-hour coculture with NK cells. Median, interquartile ranges, minima, and maxima are shown. Cells are normalized by dividing average of the 3 technical replicates for each condition from each patient by the corresponding average of CML cells–only condition for each patient. Control CD34⁺ patient cells are represented by the dashed line. P values have been calculated with a paired t test. (C) Lineplot of CD34⁺ cell viability of 2 patients at different E:T ratios, 1 (left) being highly sensitive to birinapant treatment alone, and the other (right) being resistant to birinapant treatment alone. 0 represents control, no NK cells. (D) Box plot showing drug sensitivity profiling of 2 healthy donor bone marrow CD34⁺ cells’ sensitivity to NK cells at different E:T ratios with and without drug treatment ex vivo. Dots represent normalized healthy CD34⁺ cells of the 2 donors after 24-hour coculture with NK cells. Median, interquartile ranges, minima, and maxima are shown. Cells are normalized by dividing the average of the 3 technical replicates for each condition from each donor by the corresponding average of CD34⁺ cells–only condition for each patient. Control CD34⁺ cells are represented by the dashed line. (E) Bar plot of normalized total colony counts from a colony forming assay done on 2 healthy samples and 3 CML samples. Total colony counts from each condition have been normalized to the CD34-DMSO condition within each sample. (F) Box plot showing effect of drug treatment on the efficacy of unexpanded NK cells at an E:T ratio of 2:1 from 3 patients with CML, receiving imatinib (right panel), healthy control NK cells from 3 healthy donors (middle panel), and control K562-luc cells exposed only to drug and not NK cells. Median, interquartile ranges, minima, and maxima are shown.