Fig. 2: Efficacy of BH3 domain peptides at inducing MOMP when added to permeabilized cells.

Mitochondrial assays were performed on permeabilized parental KMS-12-PE cells (A) or their isogenic BAX^-/-BAK^-/- counterparts (B) after treatment for 1.5 h with 0–160 µM of the indicated BH3 peptide; the binding specificities of peptides are shown in Fig. S2A. Identical mitochondrial assays were undertaken with permeabilized parental (C) or BAX^–/–BAK^–/– (D) AMO1 cells. (E) A peptide from the BH3 domain of BIM (BH3^BIM) causes BAX/BAK-dependent release of Cytochrome c. Permeabilized AMO1 cells of the indicated genotype were treated with 0–10 µM of the BH3^BIM peptide for 1 h, fractionated and blotted for Cytochrome c; loading control: β-actin. The blots are representative of 2 independent experiments; original uncropped immunoblots are provided with Supplemental materials. Data from treatment with other BH3 reagents is shown in Fig. S2F. F BH3^BIM peptide induces marked mitochondrial depolarization when added to permeabilized CLL cells. These permeabilized cells were treated with the indicated peptide and mitochondrial depolarization determined 1.5 h later. Data represents mean IC₅₀ ± SD of the 6 patient samples studied; the experiment was performed once per sample. The data in (A)–(D) represents the means ± SD of ≥3 independent experiments. IC₅₀ indicated in parentheses. See also Fig. S2.