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. Author manuscript; available in PMC: 2019 Sep 1.
Published in final edited form as: Clin Cancer Res. 2018 Dec 17;25(5):1574–1587. doi: 10.1158/1078-0432.CCR-18-2036

Figure 2. AZD8055 treatment selectively and significantly reduces primary CLL cell viability preferentially in poor prognostic subsets.

Figure 2

A. Primary CLL cells were incubated with increasing concentrations of AZD8055 or left untreated for 48 h (n=12 CLL patient samples). Cell viability was analyzed by flow cytometry, with the percentage of viable cells assessed as Annexin V-7-AAD-; B. Cell viability was compared upon treatment of primary CLL cells with 100 nM AZD8055 (AZD; blue), 10 nM rapamycin (RAP; red) or 1 μM ibrutinib (IB; pink) for 48 h, compared with NDC (white), (n=15); C. The level of apoptosis (Annexin V+7-AAD-) was compared upon treatment of primary CLL cells with 100 nM AZD8055, 10 nM rapamycin or 1 μM ibrutinib for 48 h, relative to NDC. p values generated by a paired two tailed t test (n=15). Inset: Western blotting was performed to show the levels of PARP cleavage (cPARP) in primary CLL cells treated with 100 nM AZD8055, 10 nM rapamycin or 1 μM ibrutinib for 48 h, or left untreated (NDC). GAPDH is included as a loading control. D. The level of apoptosis induced in primary CLL cells treated with 100 nM AZD8055 minus background (NDC) was compared between cytogenetic subgroups: good (Norm/del(13q)) vs poor (del(11q) or (17p)). p value generated by an unpaired two tailed t test (n≥7 per subgroup). E. Freshly-isolated peripheral blood mononuclear cells from healthy donors, were incubated with 100 or 300 nM AZD8055, 10 nM RAP or 1 μM IB for 48 h, compared with NDC. Cell viability of B (CD19+) and T (CD3+) populations was assessed, as indicated (n=6).