Evaluation of LQB118 inducing apoptosis on <i>L. braziliensis</i> – A) and B) Phosphatidylserine exposure on promastigotes.

<p>Promastigotes were incubated with 3,5 or 20 µM LQB118 to 24 or 48 h/28°C and then stained with Annexin V-FITC+propidium iodide and analyzed by flow cytometer. Controls were promastigotes incubated with 60 µM Miltefosine or the culture medium supplemented with 20% fetal bovine serum. In <b>A</b> only treatment at 48 h and in <b>B</b> quantitative evaluation of cells stained with annexin V-FITC at 24 and 48 h. (mean ± SD, n = 3). * P<0.05 ** P<0.01. <b>C) ROS generation and D) impairment of ATP production in LQB-118-treated promastigotes</b>. Promastigotes of <i>L. braziliensis</i> were incubated for 48 h in the presence of LQB-118 in Schneider's insect medium plus 10% HIFCS. <b>A</b>) ROS generation was quantified using H₂DCFDA, <b>B</b>) Cellular ATP concentration was measured by bioluminescence assay. Results are presented as means ± standard error; <i>n</i> = 3. *<i>P</i><0.05; **<i>P</i><0.01. <b>E) </b><b><i>In situ</i></b><b> DNA fragmentation of intracellular amastigote</b>. Infected monolayers of hamsters peritoneal macrophages were treated with the indicated concentrations of LQB118 for 48 h. Monolayers were labeled using TUNEL and observated using fluorescence microscopy. Highlight the cells in 400× magnification. Legend Arrows indicate intracellular amastigotes, N – Macrophage nucleus.