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. 2023 Jul 20;13:11737. doi: 10.1038/s41598-023-38127-4

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© The Author(s) 2023

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Effect of 2HPCD treatment on cholesterol dynamics in Ae. aegypti midguts and Ae. albopictus cell lines. (a) Ae. aegypti mosquitoes were injected with 10 mM of 2HPCD or PBS, left to recover for 2 days before a bloodmeal was given. 72 h post bloodmeal midguts were dissected, fixed and stained with Nile red (red) and DAPI (blue) to detect intracellular lipid droplets (distinct punctate staining as outlined in white boxes) and cell nuclei respectively. For each experiment 4 midguts were included and analysed, figure represents a typical image of the sets. (b) Cells were pulse labelled for 30 min with Topfluor (green), a fluorescent cholesterol derivative and treated for 48 h in either 2HPCD or PBS in order to measure cholesterol dynamics. Cell nuclei were stained with DAPI (blue). (c) Cellprofiler was used to calculate the number of Topfluor spots per cell in 3 independent replicates for each treatment.