Figure 7: Nuclear changes of saffron-treated A549 cells. A549 cells seeded on chamber slides were treated with media or different saffron concentrations. Suspension of the A549 cells was incubated with media (a), 200 μg/mL of saffron (b), and 400 μg/mL of saffron (c) for 48 hours at 37°C to induce apoptosis. Cells were then labeled with FAM-VAD-FMK for 60 minutes at 37°C. Cells were washed, and then Hoechst stain was added and incubated for 5 minutes. Wet-mount slides were prepared and the photos of the same cells were taken and superimposed. Caspase activity (green) was detected using a band pass filter (excitation at 488 nm, emission at 520 nm). Nuclear staining by Hoechst stain (blue) was revealed using a UV filter (excitation at 365 nm, emission at 480 nm). In these pictures, the yellow arrows represent the nuclear breakdown and chromatin condensation of A549 cells induced by saffron.