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Fig. 5 | Journal of Hematology & Oncology

Fig. 5

From: Targeting the CXCR4 pathway using a novel anti-CXCR4 IgG1 antibody (PF-06747143) in chronic lymphocytic leukemia

Fig. 5

PF-06747143-induced cell death is bivalency-dependent, involves ROS, but not caspase activation. a Primary leukemia B cells obtained from a CLL patient were cultured alone or in presence of stroma-NK-tert cells after treatment with 10, 100, and 1000 nM of m15-IgG1, PF-06747143, or its Fab and F(ab’)2 forms, for 48 h. % SICD was measured by flow cytometry using CD19/CD5/Annexin V staining. The results of samples analyzed in duplicates with the mean ± SD are shown for each group. b CLL cells were incubated with PF-06747143 (100 nM), F-ara-A (10 μM), or etoposide (30 μM) for 48 h, either alone or in combination with different concentrations of a pan-caspase inhibitor, Z-VAD-FMK (Z-VAD) (10, 30, 90 μM). CLL cell death was analyzed by flow cytometry. c CLL-B cells (n = 6 per group) were incubated for 48 h with the intact CXCR4 antibody PF-06747143, the Fab and the F(ab’)2 forms of the antibody, the CD20 antibodies rituximab or obinutuzumab, or the IgG1 control antibody. H2O2 was used as a positive control. ROS production and cell dealth were analyzed by co-staining for ROS and CD19+/CD5+/Annexin V, respectively. The figure shows the individual data points for each group, and horizontal lines represent the mean of each group. Statistical significance was determined using Bonferroni’s correction test where *, **, ***, and **** represent p < 0.05; p < 0.01, p < 0.001, and p < 0.0001, respectively

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