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. 2019 Apr 16;9:6114. doi: 10.1038/s41598-019-42273-z

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Evaluation of reactive oxygen species (ROS) and intracellular calcium levels in L. (L.) infantum promastigotes treated with compound 2 (190 μM). (A) H₂DCFDA dye fluorescence was measured by spectrofluorimetrically (excitation 485 nm and emission 520 nm) after 2 h of incubation. Untreated promastigotes and treated with H₂O₂ (400 μM) were used to achieve minimal and maximal depolarization, respectively. (B) Fura-2 AM dye fluorescence was measured spectrofluorimetrically (excitation 360 nm and emission 500 nm) after 5, 20, 60 and 120 min of incubation. Untreated promastigotes and treated with TX-100 (0.5%) were used to achieve minimal and maximal depolarization, respectively. Fluorescence is reported as percentage relative to untreated promastigotes (100%). A representative experiment is shown. ***p < 0.0001.