Enhanced induction of apoptosis markers in NSCLC cells by combination of AA and 3-PO.

<p>(A) Analysis of early apoptotic cells and (B) late apoptotic cells by annexin V binding and propidium idodide staining. H1299 cells were treated with either AA (300 µM), 3-PO (10 µM) or a combination of AA and 3-PO for the indicated times. Cells were then stained with annexin V and propidium iodide and visualized by fluorescent microscopy. A minimum of 200 cells were scored for each sample and the data represent means ±SEM from three independent experiments. (C) Analysis of caspase 3/7 activity. H1299 cells were treated with either AA (300 µM), 3-PO (10 µM) or a combination of AA and 3-PO for the indicated times. Caspase 3/7-positive cells were identified using a fluorogenic caspase substrate specific for activated caspase 3 and/or 7 (CellEvent™ Caspase-3/7 Green). Data represent means ±SEM from three independent experiments where 150–200 cells per sample were visualized in each experiment. a; statistically significant difference (<i>P</i><0.05) from vehicle treated control. b; statistically significant difference (<i>P</i><0.05) from control and individual AA and 3-PO treatments.