Fig. 6

High BCL-2 dependence predicts favorable drug response in primary CLL cells ex vivo. A Baseline BH3-profiling was performed on primary CLL cells. Cells were incubated in specific BH3 peptides or mimetic for 1 h prior to determining CytC loss. Patients arbitrarily labeled as patient 1–8. B Western blot analysis showing MCL-1, BCL-2, BIM xL, L, S, BAX and β-Actin levels of the same 8 CLL patient samples in Fig. 6A. C, D Pearson correlation coefficient analyses between BCL-2 family protein expressions, normalized to β-Actin levels from Fig. 6B. E Pearson correlation coefficient analyses of BCL-2 dependence and normalized BCL-2, BAX or BIM protein expression levels. F Replot of baseline BH3-profiling of primary CLL cells from patient 1 and 7 with different degrees of MCL-1 and BCL-2 dependences. G Cell death levels by fold change (F.C.) of annexin V staining of the same patient 1 and 7 following 24-h ex vivo fludarabine or ibrutinib treatment (with stroma NKtert co-culture) (Viability was measured in duplicates of the same patient, n = 2). H BH3-profiling performed following ex vivo treatment with DPI or AA for 6 h in 8 CLL patient primary cells to evaluate BCL-2 dependence level. I Cell death levels by fold change of annexin V staining of CLL patient primary cells following 2-h pre-treatment with AA or DPI followed by 24-h ibrutinib (n = 8) (with stroma NKtert co-culture). Sidak’s multiple comparisons test was used. J Western blot analysis showing S70pBCL-2, BCL-2 and β-Actin levels of 8 CLL patient primary cells following ex vivo treatment with DPI or AA for 6 h. A representative patient shown. Densitometry analysis is in supplemental figure S12C. K BH3-profiling performed following ex vivo treatment of 0.1 μM IACS-010759 for 6 h in 8 CLL patient primary cells to evaluate BCL-2 dependence level. L Cell death levels by fold change of annexin V staining of CLL patient primary cells following 2-h pre-treatment with 0.1 μM IACS-010759 followed by 24-h 2.5 nM ABT199 (VEN) (n = 8) or 1 μM ibrutinib (n = 7) (with stroma NKtert co-culture). Sidak’s multiple comparisons test was used