2.4.5. Apoptosis/Necrosis Detection by Flow Cytometry

Promastigotes were treated as described above, and then, the cells were removed from the culture, centrifuged, and washed with PBS pH 7.2 for incubation for 30 min with 10 μL of Annexin V-FITC, incubated with 10 μL of PI for 10 min, and read in a BD FACSCantoIITM flow cytometer. Data were analysed using FlowWin, software, 2.5.1 version (Turku, Finland) and GraphPad Prism 8.0 version. AMB (0.5 µg/mL) was used as a positive control to induce necrosis, and miltefosine (3 µg/mL) was used as a positive control for apoptosis.