FIG 8.

Analysis of AmB-induced apoptosis-like phenomena after inhibitor (I₁) treatment. (A) The figure depicts the cell death assay by flow cytometry in response to inhibitor (I₁) and the positive control (100 μM H₂O₂), where the differential changes in the apoptotic population are represented by the annexin V-positive but PI-negative cells in response to LdAPx inhibitor and the live population are represented by annexin V-FITC and PI-negative cells. (B) The rate of lipid peroxidation in AmB-sensitive and -resistant strains was measured by spectrofluorometer and is expressed as fluorescence units at 430 nm. (C) Metacaspase activities in the AmB-sensitive and -resistant parasites after exposure to inhibitor (I₁). (D) DNA fragmentation analysis by agarose gel electrophoresis. The DNA profiles by lane are as follows: lane 1, markers; lane 2, DNA of the resistant parasite; lane 3, DNA of the sensitive parasite; lane 4, DNA from the resistant AmB-treated strain; lane 5, DNA from the sensitive AmB-treated parasite; lane 6, DNA from the resistant parasite treated with AmB plus LdAPx inhibitor (I₁); lane 7, DNA from the sensitive parasite treated with AmB plus LdAPx inhibitor (I₁). (E) Measurement of released lactate dehydrogenase to determine the membrane integrity of AmB-sensitive and -resistant L. donovani parasites after treatment with 14 μM LdAPx inhibitor (I₁) for 4 h. All experiments were performed in triplicate, and data are expressed as the mean ± SD. Asterisks indicate significant difference: *, P < 0.05; **, P < 0.001.