Journal: Cell Death & Disease

Article Title: Therapeutic potential of BH3-mimetics and NK cell-mediated immunotherapy in T-ALL

doi: 10.1038/s41419-026-08698-x

Figure Lengend Snippet: T-ALL cell lines were exposed for 48 h to increasing concentrations (0.1, 1, 5, 10, 50, 100, 250, 500, 1000, 5000, 10,000 nM) of A venetoclax, B A1331852 C AZD4320 or D AZD5991 before analysis of cell death by propidium iodide staining and flow cytometry. Data points indicate mean values of N = 3 independent experiments in triplicates and error bars standard deviation. E Scatter plot of EC 50 values of the four BH3-mimetics with individual data points of all cell lines and medians (lines). Mann-Whitney test; * indicates significance, p < 0.05. F , G Association of the EC 50 values of AZD4320 with those of venetoclax and A1331852 in all cell lines. Spearman correlation (two-tailed); r, correlation coefficient; p, significance. H Protein extracts of T-ALL cell lines were co-incubated with anti-BIM-antibody overnight, co-immunoprecipitation and input protein extracts were subjected to western blot. Jurkat was used for IgG isotype control. Basal levels of proteins and protein complexes with BIM are shown in six T-ALL cell lines (representative of N = 3). I For BH3-profiling, cell lines were incubated with BH3-peptides before fixing and cytochrome c staining. Cytochrome c release after peptide exposure was analyzed by flow cytometry and values were normalized to alamethicin as positive control and DMSO as negative control. Means of N = 3 independent experiments in triplicates. BAD–HRK is the calculation of BCL-2 dependency by subtracting HRK priming (which targets only BCL-XL) from BAD priming (which targets both BCL-2 and BCL-XL). The lower heatmap shows the EC 50 values of the inhibitors indicated.

Article Snippet: To target anti-apoptotic BCL-2 family proteins, BCL-2 homology 3 (BH3)-mimetics were developed, which functionally mimic sensitizer proteins by binding to and antagonizing anti-apoptotic BCL-2 family members [ , ].

Techniques: Staining, Flow Cytometry, Standard Deviation, MANN-WHITNEY, Two Tailed Test, Incubation, Immunoprecipitation, Western Blot, Control, Positive Control, Negative Control

Journal: Cell Death & Disease

Article Title: Therapeutic potential of BH3-mimetics and NK cell-mediated immunotherapy in T-ALL

doi: 10.1038/s41419-026-08698-x

Figure Lengend Snippet: A Graphical scheme of dynamic BH3-profiling. Cell lines were exposed to drug or DMSO for 4 h (Loucy) or 2 h (other cell lines) before permeabilization and peptide exposure, fixation and cytochrome c staining. Cytochrome c release was analyzed using flow cytometry. Values were normalized to alamethicin as positive control and to DMSO as negative control. N = 3 independent experiments in triplicates. BAD–HRK is the calculation of BCL-2 dependency by subtracting HRK priming (which targets only BCL-XL) from BAD priming (which targets both BCL-2 and BCL-XL). Created with BioRender.com. B Delta-priming was determined after incubation with AZD4320 (left) and AZD5991 (right) and calculated by subtracting normalized cytochrome c release of DMSO-treated cells from normalized cytochrome c release of drug-treated cells. C ALL-SIL cells were treated with 50 nM AZD4320 and/or 1 µM AZD5991 for 4 h. After treatment, protein extraction was performed and protein lysates were subjected to co-immunoprecipitation with anti-BIM-antibody. Co-immunoprecipitation and input protein extracts were subjected to western blot analyses. Representative of N = 3 different cell lines. The graphical scheme illustrates the mechanism of action of the BH3-mimetics used for the treatment before co-immunoprecipitation. AZD4320 and AZD5991 bind to their target proteins, releasing BIM from them, which subsequently activates apoptosis. Created with BioRender.com.

Article Snippet: To target anti-apoptotic BCL-2 family proteins, BCL-2 homology 3 (BH3)-mimetics were developed, which functionally mimic sensitizer proteins by binding to and antagonizing anti-apoptotic BCL-2 family members [ , ].

Techniques: Staining, Flow Cytometry, Positive Control, Negative Control, Incubation, Protein Extraction, Immunoprecipitation, Western Blot