Evaluation of apoptosis in canine lung cancer cell lines treated with STA-1474.

<p>Apoptosis was assessed by annexin V/PI staining flow cytometry and detection of caspase 3/7 enzymatic activity. (A) Cells were treated with dimethyl sulfoxyide (DMSO, control) or 0.005–1 μM of STA-1474 for 24 h. Staining with annexin V and the vital dye, propidium iodide (PI), were used to evaluate early and late apoptosis. Cells that are considered viable are both annexin V and PI negative, while cells that are in early apoptosis are annexin V positive and PI negative, and cells that are in late apoptosis or already dead are both annexin V and PI positive. (B) Both cell lines were treated as above and evaluated for executioner caspase-mediated apoptosis. Activated caspases 3 and 7 were assessed 24 and 48 h after treatment. Experiments were performed in triplicate and repeated three times. Each graph shows mean ± SEM. *, <i>p</i><0.05; **, <i>p</i><0.01; ***, <i>p</i><0.001.