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. Author manuscript; available in PMC: 2024 Apr 17.
Published in final edited form as: Sci Transl Med. 2023 Apr 5;15(690):eadd5318. doi: 10.1126/scitranslmed.add5318

Fig. 1. Bcl-2 inhibition affects peripheral lymphocyte and bone marrow cell frequencies in vitro and in vivo.

Fig. 1.

(A) Analysis for active caspase-3/7 after Bcl-2i treatment. Peripheral blood cells and bone marrow cells were cultured with various concentrations of Bcl-2i for 24 hours (n = 4, peripheral blood; n = 3, bone marrow; each sample was obtained from different NHPs). Dose-dependent activation of caspase-3/7 was measured by flow cytometry in peripheral blood–derived CD4+ T cells, CD8+ T cells, B cells (CD3−CD20+), NK cells (CD3−CD16+NKG2a+), granulocytes, and monocytes. Dose-dependent activation of caspase-3/7 was also measured in HSCs (CD34+CD90+CD45RA−) and MPPs (CD34+CD90−CD45RA−) isolated from bone marrow. (B) Analysis for annexin V binding and PI incorporation after Bcl-2i treatment. Peripheral blood cells and bone marrow cells were evaluated for apoptosis (annexin+PI−) and death (annexin+PI+) over 24 hours in culture with Bcl-2i by flow cytometry (n = 4, peripheral blood; n = 3, bone marrow; each sample was obtained from different NHPs). (C and D) Three cynomolgus monkeys were treated with Bcl-2i (10 mg/kg) for 5 days. (C) Peripheral blood cell counts were measured at the indicated time points. (D) Bone marrow aspiration was performed before and 5 days after Bcl-2i treatment, and absolute counts of HSCs, MPPs, and CFUs were evaluated. Each value represents cell counts on day 5 relative to pretreatment values (%) (n = 5, each sample was obtained from different NHPs). Data are presented as means ± SE. Data were analyzed using one-way analysis of variance (ANOVA) for multiple comparisons. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001.