Extended Data Fig. 9: Comparison of BCL2 SMPC and PBA PPI profiling and different methods to predict responses to BH3 mimetics.
a, Schematic for BH3 profiling based on JC-1 staining, and measurement of BCL2 levels using flow cytometry or western blotting for primary AML samples. b,c, BH3 profiling on HL60 cells. (b) Relative fluorescence units (RFU) of 590 nm wavelength for HL60 cells through the treatment of BH3 peptides or BH3 mimetics, (c) Depolarization of HL60 cells through the BH3 profiling. Depolarization by 10 μM of BAD peptides was indicated. d,e, Correlations for ex vivo ABT-199 AUC with (d) depolarizations by BAD (10 μM) – HRK (10 μM) peptides, (e) combination of multiple PPI metrics (BCL2-BIMBH3 PBA, BCL2-BAX CPX, and BCLxL-BAK CPX). The outliers identified in model (d) were indicated (n = 14) (One-sided F-test, p-value = 0.011, 8.1e-05). f,g, ROC curves for ex vivo ABT-199 responses with (f) depolarizations by BAD (10 μM) – HRK (10 μM) peptides, (g) Combination of multiple PPI metrics (Two-sided t-test, p-value = 0.07, 0.009). h, Correlations between BCL2 protein levels determined by SMPC and flow cytometry for primary AML samples (n = 14). i, Prediction models for ABT-199 ex vivo AUC with BCL2 protein levels determined by flow cytometry (One-sided F-test, p-value = 0.077). j, Correlations between BCL2 protein levels determined by SMPC and western blotting for primary AML samples (n = 32). The raw blot image is provided as a Source Data. k, Prediction models for ABT-199 ex vivo AUC with BCL2 protein levels determined by western blotting (One-sided F-test, p-value = 0.0003).
