Fig. 1

Healthy hematopoiesis is affected by higher dosage combination therapy. a, b Bone marrow mononuclear cells (BMMNCs) from healthy, elderly donors were treated for 72 h with the indicated concentrations of venetoclax (VEN) and 5-azacitidine (5-AZA), alone or in combination. The viability of bulk bone marrow cells (a) (n = 7) or purified CD34⁺ cells (b) (n = 7) was measured by flow cytometry using Annexin V and 7AAD staining. Data are presented as mean ± standard deviation (SD) of the ratio between viable cells after a 72 h treatment with drug or vehicle (DMSO). One-way ANOVA resulted in p < 0.0001 for total BMMNC (a) and p < 0.0001 for the CD34⁺ compartment. Results from post hoc pairwise comparison are reported in the figure. c BMMNCs (1 × 10⁴) from 9 individual healthy donors were plated in methylcellulose after 72 h of treatment with 1 µM venetoclax and 5-AZA (1 µM, 5 µM, 7.5 µM or 10 µM as indicated), alone or in combination. The total number of colonies, distinguishing between colony-forming units (CFU) of the multi-potential granulocytic–erythroid–macrophagic–megakaryocytic lineage (CFU-GEMM), the granulocytic–macrophagic lineage (CFU-GM), and the burst-forming units-erythroid lineage (BFU-E) were determined at day 14. Experiments were performed in duplicates. One-way ANOVA resulted in p < 0.0001. The results from post hoc pairwise comparison are reported in figure