Figure 5

Hmxbato induced apoptosis in lung tumor cells. Tumor (A549) and normal (BEAS-2B) lung cells were incubated for 24 h in the absence (control) or presence of hmxbato (3.8 µM and 19 µM) and subjected to annexin V-FITC and propidium iodide double-staining assay. (A) Dot plot graphs, referring to one of the independent experiments performed, showing the distribution of A549 and BEAS-2B cells in different quadrants, according to the incorporation or not of Annexin V (AV) FITC/propidium iodide (PI). Lower left quadrant, viable cells; right lower quadrant [AV(+)], initial apoptosis; right upper quadrant [AV/PI(+)], late apoptosis; upper left quadrant [PI(+)], necrosis. (B) Bar graphs showing the quantification of A549 and BEAS-2B cells in apoptosis/necrosis. Data were expressed as mean ± standard deviation of experiments performed in triplicate. Three independent experiments were performed and over 10,000 cells were analyzed in each replicate. Statistical differences were determined using one-way ANOVA and Tukey's multiple comparison test. Significant differences with respect to control were considered when p < 0.1 (*), p < 0.01 (**), p < 0.001 (***) and p < 0.0001 (****).