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. 2018 Nov 12;8:16678. doi: 10.1038/s41598-018-34891-w

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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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COX-2 interacts with IRE1α in vivo. (A) Immunoprecipitation (IP) of endogenous IRE1α from HEK293 cells with anti-IRE1α or anti-COX-2 -antibodies. Immunoblot (IB) analysis was carried out with anti-IRE1α or anti-COX-2 antibodies. The location of IRE1α and COX-2 are indicated by the arrows. Immunoprecipitation experiments were performed in triplicate with representative blots shown. (B) His-tagged ER luminal domain of IRE1α (IRE1-NLD) was expressed in COS-1 cells followed by immunoprecipitation with anti-COX-2, anti-His-tag antibodies or IgG. Immunoblot (IB) analysis was carried out with anti-His antibodies. The location of IRE1-NLD is indicated by the arrow. Immunoprecipitation experiments were performed in triplicate with representative blot shown. (C) Pull-down of COX-2 in COS-1 cells expressing His-tagged IRE1-NLD. Upper blot was probed with anti-COX-2 antibodies. The lower blot was probed with anti-His-tag antibodies. Protein samples were separated on the same SDS-PAGE, the middle empty lanes were removed from the lower blot for the sake of clarity. Pull-down assay was performed in triplicate. The images of (A–C) shown are cropped. The full-length gels/blots are shown in Suppl. Figs S12 and S13.