Apoptotic and clonogenic effect of DhL on D384 cells.

<p>Apoptotic D384 cells were detected and monitored by Annexin V-FITC/PI staining. (A) A representative histogram is shown for each DhL concentration. Quadrant 1, PI (+) (cells undergoing necrosis); Quadrant 2, Annexin V-FITC (+) and PI (+) (cells in the late apoptosis and secondary necrosis); Quadrant 3, Annexin V-FITC (−) and PI (−) (living cells); Quadrant 4, Annexin V-FITC (+) and PI (−) (cells in the early apoptosis). (B) Apoptotic changes. The fractions of live (black bars), early apoptotic (gray bars), late apoptotic/secondary necrotic (white bars) and necrotic (striped bars) cells are shown. Symbols denote statistically significant differences: * p<0.01, ** p<0.001 and *** p<0.0001 with respect to control conditions for live cells, +p< 0.01, ++p< 0.001 and +++p< 0.0001 with respect to control conditions for early apoptosis, and ⊕ p<0.01 with respect to control conditions for late apoptosis/secondary necrosis. (C) Clonogenic assay. Cells were treated with DhL for 16 h. The number of counted colonies was expressed as a fraction of the control (defined as 100%). Symbols denote statistically significant differences: * p< 0.01, ** p<0.001 vs. control. Data are represented as the mean ± SEM (n = 4) of 3 independent experiments.