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. 2020 Jul 8;10:11207. doi: 10.1038/s41598-020-67466-9

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© The Author(s) 2020

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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Dafachronic acid accelerates the complete molting of infective third stage D. immitis larvae to L4. The iL3 stage larvae were collected from mosquitos and treated with Δ7- (Δ7-DA, A and B) or Δ4-dafachronic acid (Δ4-DA, C and D) at 10 µM or DMSO. The developmental changes were observed daily under a microscope. The iL3 that completed the molting from iL3 to L4 (left panel A and C) were scored as well as the iL3 that were still undergoing molting (right panel B and D). The number of free cuticles (left panel) found in the wells are directly related to the amount of larvae that had completed the molt and were L4. The figure represents the mean of two independent experiments performed in quadruplate from different pools of iL3s using Prism 6.0 (Graph Pad Software, Inc.). The statistical significance was evaluated with an unpaired t test (**p < 0.005; *p < 0.05).