Figure 2
Analysis apoptotic cells with SFN treatment. A. Apoptotic cells were analyzed by using Annexin V FITC apoptosis detection kit (Calbiochem, Gibbstown, NJ). Control and SFN treated MDAH-2774 cells (1 × 105 cells) were fixed, stained both with Annexin V FITC and with propidium iodide (20 μg/ml) and were processed for flow cytometric analysis. B. Cells were mixed with annexin V-biotin, incubated for 15 min at room temperature, and treated with streptavidin conjugated to FITC. Apoptotic cells within the same microscopic field were viewed and photographed by phase contrast and by fluorescence. Using the FITC filter, early apoptotic cells (positive for Annexin V-Biotin-FITC staining) appear bright green. C. Western blot analysis of poly (ADP-ribose) polymerase (PARP) protein, caspase 9, BAK, Bcl-2 and control β actin with increased doses of SFN. Native and cleaved PARP, pro and active caspases are indicated by arrows.