Fig. 2: Tracking the PPI profile changes to intrinsic apoptotic stress and assessment of potential PPIs with PBA.
a–d, Changes in endogenous BCL2-family PPI profiles of HL60 cells through apoptosis progression by 2 μM of staurosporine. Active BAX/BAK level (a), total levels of anti-apoptotic proteins (b), BIM complexes (c), BAX complexes (n = 10 independent images) (d). e, Schematic of the PBA to measure the unoccupied populations of surface-immobilized anti-apoptotic proteins. f, Changes in BIMBH3 PBAs for anti-apoptotic proteins from HL60 cells through apoptosis progression by staurosporine (n = 10 independent images). g, Schematic of the comparison of the BCL2 protein complex compositions in different AML cell lines (left). The density of surface-immobilized BCL2 was constantly maintained by optimizing the total protein concentration of crude cell extracts in all AML cell lines (right) (n = 10 independent images). h–j, Compositions of the BCL2 PPI profiles in AML cell lines. BCL2 protein complexes (h), BCL2-BIMBH3 PBA (n = 10 independent images) (i), integrated BCL2 complex compositions (j). k, Changes in BCL2-BIMBH3 PBA and BCL2-BAD PBA with increasing amounts of BAD probe (n = 10 independent images). BIMBH3 probe was presented at 10 nM. The single-molecule counts were rescaled to account for the labelling efficiencies of the immunoassays calculated in Extended Data Fig. 5. Data represent means ± s.d.
