FIGURE 3 from Identification and Characterization of a Small Molecule Bcl-2 Functional Converter

BFC1108 activates intrinsic mitochondrial death pathway. A, BFC1108 suppresses the colony-forming ability of MEF cells in a Bcl-2–dependent manner. WT and Bcl-2−/− MEF cells were treated for 48 hours in a medium containing 10% FBS and the colony formation was determined after 2 weeks. B, Quantification of colony formation data shown in A. **, P < 0.01. C, Bax or Bak is required for BFC1108-induced suppression of viability. WT MEF, Bax Knockout (Bax−/− Bak+/+), Bak knockout (Bax+/+ Bak−/−) and double knockout (Bax−/− Bak−/−) MEF cells were treated with 1 µmol/L BFC1108 for 24 hours in 10% FBS medium and viability was assessed using CellTiter-Glo assay. **, P < 0.01; ***, P < 0.001; ns, not significant. D, BFC1108 decreases mitochondrial membrane potential of Bcl-2–expressing H460 lung cancer cells. JC-1 dye was used to stain live H460 cells that were treated with 10 µmol/L BFC1108 for 16 hours in 10% FBS containing medium. Images taken with FITC, and rhodamine filters were overlaid. Cells stained orange have intact mitochondrial outer membrane and the ones turning green have compromised outer membrane indicating loss of membrane potential.