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. 2017 Nov 28;7:16497. doi: 10.1038/s41598-017-16662-1

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© The Author(s) 2017

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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mTOR-dependent IRE1 attenuation of RNAse activity has a significant impact on ER remodelling dynamics. (A) A quantitative imaging approach to estimate the extent of ER remodelling in human MCF10A epithelial cells. Main image processing and analysis steps and segmentation examples are shown in the schematic depiction. (B and C) A measurement of relative ER density distribution comparing inner and peripheral regions of the cell captures ER expansion in tunicamycin-treated MCF10A cells, as compared with mock-treated cells. Four experimental replicates, containing ~1200 cells each, were imaged and quantified as indicated. Wash-out of the ER stressor leads to reaccomodation of the ER towards perinuclear regions over time [C; lower left panel and graph]. mTOR inhibition immediately after ER stressor washout however delays this reaccomodation. Lower panel row shows magnified images of the selected ROIs.