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. 2024 Mar 15;43(8):1653–1685. doi: 10.1038/s44318-024-00063-y

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© The Author(s) 2024

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 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/. Creative Commons Public Domain Dedication waiver http://creativecommons.org/publicdomain/zero/1.0/ applies to the data associated with this article, unless otherwise stated in a credit line to the data, but does not extend to the graphical or creative elements of illustrations, charts, or figures. This waiver removes legal barriers to the re-use and mining of research data. According to standard scholarly practice, it is recommended to provide appropriate citation and attribution whenever technically possible.

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Figure EV5 — (A) Immunoblot analysis of the indicated organellar markers in whole-cell lysates (lysate), crude membranes (P100), and MemPrep isolates (isolate). ER membranes were immuno-isolated via the Rtn1-bait protein. Sec61 and Dpm1 are prototypical ER membrane markers. Por1 is a marker for the outer mitochondrial membrane, Vph1 is a vacuolar marker. Pep12 marks endosomes and Gas1 serves as plasma membrane marker. 1 µg total protein loaded per lane. (B) Quantification of the organelle markers Dpm1, Vph1, Por1, Pep12 and the Rtn1-bait protein from three immunoblots of independent replicate ER MemPreps after prolonged proteotoxic stress induced by DTT (n = 3 biological replicates). (C) Quantification of three immunoblots from independent replicate ER MemPreps after prolonged proteotoxic stress induced by TM. Error bars indicate standard deviations (n = 3 biological replicates). (D) Correlation of DTT- and TM-induced fold changes, after Limma analysis, over pre-stress with a Pearson correlation coefficient r = 0.82. K-means clusters are indicated by colored groups and their respective cluster number (n = 3 biological replicates). (E) Gene ontology term enrichments in K-means clusters (n = 3 biological replicates). Data information: Data in (B, C), data from three biological replicates are presented as individual data points and as the mean ± SD. Data in (E) from n = 3 biological replicates are presented as the mean. P values were derived from a Fisher-test and corrected for multiple testing with the method of Benjamini and Hochberg. Source data are available online for this figure.