Fig. 2

Dual treatment with FCCP and TRAIL induces apoptosis and disrupts cellular metabolism. Dual treated cells were analyzed for extracellular expression of phosphatidylserine by antibody labeling with Annexin V (AnnexinV:FITC), the fluorescent metabolic dye resazurin (Resazurin:PE), and viability probe DAPI to determine apoptotic induction. a Co-labeling with AnnexinV:FITC and DAPI indicated high viability in control wells, with significant increases in both apoptotic cells (DAPI⁺FITC⁺; purple) as well as nonviable cells (DAPI⁺FITC⁻; blue) following both FCCP and TRAIL (c: n = 3 ANOVA p < 0.001**). b The increase in DAPI⁺ events was also present when evaluating changes in global cellular metabolism, indicative of both respiring non-viable cells, as well as non-respiring non-viable cells. Additionally, there was a significant decrease in viable and metabolically active cells (DAPI⁻PE⁺; green) only after dual treatment with FCCP and TRAIL (d: n = 3 ANOVA p < 0.001**.) e In Kuramochi cultures, there was significant increases in both apoptotic cells as well as nonviable cells following treatment with both FCCP and TRAIL in the Kuramochi cell line (n = 3 ANOVA p < 0.001**), as well as (f) nonviable respiring cells (n = 3, ANOVA p < 0.001**). g, h SKOV3 cultures had no significant changes in apoptotic induction or metabolic modulation following treatment with FCCP and TRAIL