Functional BH3 profiling and paired cell-death testing
Cell-based study · Murine leukemia model lines dependent on human BCL-2, BCL-XL, MCL-1 or BFL-1, plus BAX/BAK double-knockout control cells; permeabilized-cell mitochondrial and intact-cell assays. [1]
Gossypol showed no mitochondrial activity at concentrations up to 10 µM in this functional profiling system, despite intact-cell killing after 24 h. The authors also observed some killing of BAX/BAK double-knockout cells. These findings limit its use as a potent direct BH3 mimetic in these models; they do not measure purified-protein binding or establish the absence of all indirect BCL-2-family effects.
| Endpoint | Reported result | Reference |
|---|---|---|
| Direct mitochondrial cytochrome c release after 1 h | Functional mitochondrial response No mitochondrial activity at concentrations up to 10 µM | [1] |
| Intact-cell death after 24 h | Annexin V positivity Cell killing occurred, including a lesser degree of killing in BAX/BAK double-knockout cells; no exact percentage or IC50 extracted. | [1] |
Literature-Reported Treatment Conditions
- Treatment concentration
- Mitochondrial activity assessed up to 10 µM, as stated in Discussion; intact-cell concentration series shown in Figure 2A, without extracting exact individual doses from the log plot.
- Exposure duration
- 1 h for iBH3 mitochondrial response; 24 h for intact-cell death
- Method
- Direct exposure of digitonin-permeabilized cells for mitochondrial profiling; separate intact-cell compound exposure for viability testing.
- Vehicle
- DMSO stock solvent; final DMSO percentage not stated in the read assay methods.
- Co-treatment
- Gossypol alone; other compounds were comparator treatments, not combinations.
- Reported limits
- Gossypol catalog number, enantiomer ratio and lot identity are not specified in Compounds. Exact individual concentration steps are not enumerated in the read text.
Methods and source locations
Compound reported in the paper: Gossypol; Compound label in the paper: Sigma-Aldrich gossypol; stereochemistry not specified
Applies to the reported Sigma-Aldrich gossypol and the specified functional assays; it does not validate an Adooq lot or establish universal target selectivity.
Figure 2A and caption; Figure 1B validation; Materials and methods: Compounds, iBH3 profiling, Viability testing, Data reduction; Results gossypol paragraph; Discussion
Direct mitochondrial cytochrome c release after 1 h: iBH3 flow cytometry in permeabilized cells; Figure 2 caption specifies N ≥ 6 and SD bars. Buffer-only and no-antibody controls define signal range; BH3 peptide responses and alamethicin validate the model panel. — Figure 2A and caption; Figure 1B validation; Materials and methods: Compounds, iBH3 profiling, Viability testing, Data reduction; Results gossypol paragraph; Discussion
Intact-cell death after 24 h: Annexin V/Hoechst flow cytometry; Figure 2 caption specifies N ≥ 3 and SD bars. Paired mechanistic comparison with iBH3; no curve-derived decimals. — Figure 2A right panel and caption; Results gossypol paragraph; Viability testing