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. 2016 Oct 12;6:35067. doi: 10.1038/srep35067

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Copyright © 2016, The Author(s)

This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

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(A) The influence of PMA on ADAM17 in HEK293 cells overexpressing iRhom1 was analysed. Therefore, cells were transfected either with an empty vector (vec) or with iRhom1_M6P (iR1). After 48 hours cells were left unstimulated (unstim) or were stimulated with 100 nM PMA for 5 minutes. 8 hours or 24 hours after PMA stimulation the cells were harvested. Glycosylated proteins were enriched by precipitation with ConA-Sepharose and immunoblotted. (B,C) Amount of mature ADAM17 and its proform was measured by densitometry of immunoblots (A). The results are mean values of three independent experiments. (B) Shown are the ratios of mature ADAM17 to the total amount of ADAM17 normalised to unstimulated and untransfected cells (vec/0 h). (C) Comparison of the rate of downregulation and recovery. Each setup (vector and iRhom1 overexpression) was normalised to its initial amount of mature ADAM17 (0 h).