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. 2018 Aug 22;9:3359. doi: 10.1038/s41467-018-05554-1

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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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Fig. 5 — A subset of class 1 mutations specifically disrupts PHD2-mediated hydroxylation. a, b Biotinylated HIF-2α peptides (residues 523–542) were immobilized on streptavidin- agarose beads and incubated with HIS₆-PHD2 for 1 h at room temperature prior to incubation with in vitro transcribed and translated (IVTT) pVHL. Streptavidin beads were pulled down (PD) and levels of HA-tagged pVHL were visualized via immmunoblotting (IB). c HEK293a cells were co-transfected with FLAG-VHL and HA-HIF-2α constructs. FLAG was immunoprecipitated (IP) and levels of HA-tagged HIF-2α were visualized via immunoblotting (IB). * denotes uncharacterized band