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. 2020 Jan 21;10:898. doi: 10.1038/s41598-020-57652-0

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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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Time-lapse endocytosis of AF747-FSA in the presence or absence of monensin and LysoTracker. LSEC cultures were preincubated with LysoTracker (yellow) and challenged with AF647-FSA (magenta) in the absence or presence of monensin (10 µM). The uptake was monitored in real-time by DV overnight. Colocalization values for FSA and LysoTracker (shown in merged images) are Pearson’s correlation coefficients calculated using the Costes threshold in Volocity Quantitation software. Images represent maximum intensity projections of 4–9 µm 3D z-stacks. Right panel shows randomly selected line profiles within the cells, indicating the degree of colocalization of the two probes. The results are representative of experiments performed with cells isolated from 4 animals, and included 7 culture dishes, 49 fields of view, and approximately 350 cells; of which 2 animals, 2 culture dishes, 17 fields of view, and approximately 100 cells were in the presence of monensin).