Figure 4

Morphology changes and DNA fragmentation in ovarian cancer cells after HNTMB treatment. (A) Morphological changes following HNTMB treatment. SKOV-3 cells were treated for 24 h with 0.2 μM HNTMB, fixed and stained with 4'-6-Diamidino-2-Phenylindole (DAPI) as described (Materials and Methods) before mounting. Microscopy was carried out (Nikon Eclipse TE2000-E inverted microscope, 20× objective) and representative images were taken. Bar = 10 μm. (B) TUNEL assay. SKOV-3 cells were treated with 0.2 μM of HNTMB for 24 h. Labeling of DNA nicks with fluorescein-12-dUTP and chromatin counterstaining with propidium iodide was carried out as described (Materials and Methods). Representative images were taken, apoptotic stain (FL-dUTP, green) and nuclear stain (Pi, red) overlaid; TUNEL positive nuclei due to DNA fragmentation appear as yellow areas. Bar = 10 μm.