Compound-associated cell death is time-dependent.

<p><i>L</i>. <i>major</i> promastigotes were treated with solvent control (1% DMSO), apoptosis inducer miltefosine (122.7 μM), compound 1 (100 μM), and 2 (100 μM) in a time course experiment for 6 h, 10 h, and 24 h. Cells were harvested, stained with AV and PI and subsequently analyzed by flow cytometry. Analysis for compound 2 was performed in an independent experiment. Percentage for each quadrant is written below the corresponding dot blot figure. Lower-left (LL): live cells (AV^-/PI^-); upper-left (UL): early necrotic cells (AV^-/PI⁺); upper-right (UR): necrotic/late apoptotic cells (AV⁺/PI⁺); and lower-right (LR): early apoptotic cells (AV⁺/PI^-).