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. 2019 Jan 24;9:438. doi: 10.1038/s41598-018-36944-6

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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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Validation and optimization of the L. donovani assay for compound screening. (A) Heatmaps of the distribution of BMDM according to the amount of intracellular amastigotes for each experimental condition, for different macrophage seeding densities (20,000 or 40,000 cells per well), MOIs (1:3, 1:6 or 1:9 macrophage to parasite ratio), and treatments (mock [Ø], DMSO and Amphotericin B [AmB]). (B) Boxplots of percentages of BMDMs normalized to mock control values obtained with 20,000 and 40,000 seeded macrophages and recovered after three days of treatment with DMSO (vehicle control), AmB (positive control) and cycloheximide (Cyclo, toxicity control) are represented. (C) Boxplots of percentages of BMDMs harboring parasites after 3 days for all experimental conditions except for cycloheximide. Whisker plot represents median and 5^th/95^th percentiles.