Dleu7 expression results in cell death. (A) A549 cells were infected with 80 MOI of Ad5-DLEU7 or Ad5-control virus; 2 days later, cells were harvested and used in the Click-iT EdU Flow Cytometry Assay. Plot data for cells labeled with Click-iT EdU Pacific Blue fluorescence (y-axis) and cells labeled with Click-iT CellCycle 488-Red (7-ADD) fluorescence (x-axis). A total of 3 subgroups, representing the percentage of cells in S phase, G₁ phase, and G₂ phase of cell cycle, are shown for cells infected with Ad5-DLEU7 and Ad5-control, respectively. (B-C) A549 cells were infected with 350 MOI of Ad5-DLEU7 or Ad5-control virus; 4 days (B) or 5 days (C) later, cells were harvested and treated with APC annexin V conjugate, which binds apoptotic cells. Panels show plot data for cells labeled with APC annexin V fluorescence (y-axis) and GFP green fluorescence for infected cells (x-axis). A total of 2 subgroups, representing the percentage of apoptotic cells and viable cells, are shown. (D-E) A549 cells were infected with 350 MOI of Ad5-DLEU7 or Ad5-control virus; 4 days (D) or 5 days (E) later, cells were harvested and treated with MitoProbe DiIC₁(5) dye, which accumulates in mitochondria with active membrane potentials. DiIC₁(5) stain intensity decreases in apoptotic cells. Panels show plot data for cells labeled with MitoProbe DiIC₁(5) fluorescence (y-axis) and GFP green fluorescence for infected cells (x-axis). A total of 2 subgroups, representing the percentage of apoptotic cells and viable cells, are shown.