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. 2021 Aug 3;18:36. doi: 10.1186/s12987-021-00266-z

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© The Author(s) 2021

Open AccessThis 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/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 1 — Subcellular localization of Pgp in RBE4- and hCMEC/D3-MDR1-EGFP cultures and endolysosomal staining. MDR1-EGFP transduced RBE4 (A) and hCMEC/D3 (B) cells grown on glass coverslips were treated with the fluorescent acidotropic LysoTracker probe to stain endolysosomal vesicles. Pgp localization visualized by green fluorescence of the EGFP protein tag and colocalization with endolysosomal vesicles was assessed by live-cell imaging and confocal laser scanning microscopy. Imaging revealed that, as expected, Pgp-EGFP (green) was localized at the cell surface in both cell lines (arrows). In hCMEC/D3-MDR1-EGFP cells, Pgp-EGFP (green) colocalized with LysoTracker (blue) stained intracellular vesicles (boxes in B), whereas this was hardly observed in RBE4-MDR1-EGFP cells. Boxes in the bottom right corner of the merged images show magnifications of Pgp subcellular localization in the respective cell line