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. 2020 Mar 24;10:5356. doi: 10.1038/s41598-020-62202-9

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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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Large CD151 patches coincide with intracellular actin accumulations. CD151-GFP transfected HaCaT cells were treated for 3 h without or with PsVs. Membrane sheets were generated, stained, and imaged by confocal microscopy. Green (CD151-GFP; GFP signal was enhanced by nanobodies), red (PsVs visualized by L1 antibody labeling) and cyan (filamentous actin; fluorescently labelled phalloidin). Images are displayed using a linear lookup table. For each channel, the same arbitrary scaling was applied. (A) For each condition a membrane sheet is shown. Magnified views from the white boxes are shown, illustrating the individual channels. (B) For the CD151 and the actin channels, within a freehand ROI excluding membrane edges, the pixels with an intensity higher than the average ROI intensity were selected. Then, the number of pixels positive in both channels were related to the number of all positive pixels in the CD151 channel. Values are given as means ± SD (n = 45 membrane sheets collected from three biological replicates). Unpaired Student’s t-test (***p < 0.001).