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. 2018 Oct 8;8:14966. doi: 10.1038/s41598-018-33378-y

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© The Author(s) 2018

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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Study on the reversibility of selected agents on protein secretion inhibition. U2OS cells coexpressing streptavidin-KDEL and ss-SBP-GFP were treated with selected agents at selected concentrations (20 μM for cyclosporin A, fendiline, parbendazole, paroxetine, parthenolide, quinacrine, sertraline, spiperone, thimerosal; 10 μM for astemizole and perhexiline) for 4 h. Then the drugs were washed out and replaced with fresh medium for cells to recover from the treatment. After different recovery periods, cells were incubated with biotin for another 4 h before fixation and image acquisition (A). Representative images of BFA (which is relatively irreversible) pretreated cells as well as parbendazole (which is highly reversible) treated cells after different recovery periods are reported in (B), scale bar equals 10 μm. Quantitative data was normalized to untreated controls and summarized as heat map, each block represents the mean value of 4 repeated measurements (C). Statistical analysis was performed by means of multiple t test, *p < 0.001 as compared to untreated controls.