A representative FACS profile of cell size (A), DHE fluorescence (B) and apoptotic cells (C) between anti-CD3 plus anti-CD28-stimulated and medium-treated CD4+CD25- T cells is displayed. A. Profile of FSC vs SSC is displayed to show the cell size. The cells were electronically gated as two populations based on their size. R1 (red) represents live or early apoptotic cells (see C). R2 (green) represents dead and/or late apoptotic cells (see C). B. Profile of DHE fluorescence (ROS+) on FL-2 vs. FSC of R1 and R2 cells. The values are shown as the MFI of R1 and R2 cells (R1/R2). Data not shown here are the MFI of unlabeled cells (negative control for DHE staining) on FL2, which is usually < 10. C. The profile of Annexin-V vs. 7-AAD staining of cultured cells compensating the R1 and R2 regions as gated in A. The quadrant gates were set according to the negative isotype control antibodies in the respective cells.