Accumulation of poly (adenosine diphosphate-ribose) by sustained supply of calcium inducing mitochondrial stress in pancreatic cancer cells

Figure 3 Confirmation of the expression of apoptosis-inducing factor followed by an increase in apoptosis in pancreatic cancer cells (AsPC-1 and CFPAC-1). A: Immunocytochemical staining for apoptosis-inducing factor (AIF) release from mitochondria and translocation into the nucleus. Red dye: AIF; Green dye: Mitochondrial indicator. Scale bars: 10 μm; B: Quantitative analysis of the mean fluorescence intensity (MFI) of AIF in CFPAC-1; C: Quantitative analysis of the MFI of AIF in AsPC-1; D: Fluorescent microphotographs for colorimetric terminal deoxynucleotidyl transferase -mediated dUTP nick end labeling (TUNEL) detection. Green dye: TUNEL staining. Scale bars: 50 μm; E: Quantitative analysis of the % ratio of TUNEL detected cells per total pancreatic cancer cells; F: Representative flow cytometry plots using Annexin V-fluorescein-5-isothiocyanate/propidium iodide staining for apoptosis; G: Representative microphotographs to compare cell condition after calcium supply; H: Quantitative analysis of the percent cell viability following calcium supply. The cells were treated with 2.5 mM lactate calcium salt for 72 h. Results represent the mean ± SD. ^aP < 0.001 vs untreated. CaLac: Lactate calcium salt; AIF: Apoptosis-inducing factor; MFI: Mean fluorescence intensity; TUNEL: Transferase-mediated dUTP nick end labeling.