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. 2016 Nov 8;2:16040. doi: 10.1038/celldisc.2016.40

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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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Degradation of unassembled soluble proteins requires p97 and Npl4. (a) A p97 dominant negative mutant inhibits the degradation of unassembled FNTA. HEK293T cells transfected with FNTA-FLAG together with the indicated plasmids were subject to cycloheximide chase analysis. (b) Knockdown of p97 stabilized FNTA. HEK293T cells transfected with FNTA-FLAG together with the indicated siRNA were analyzed by cycloheximide chase. (c) Interaction of p97 with FNTA. Cells transfected with the indicated plasmids and siRNAs were subject to immunoprecipitation and immunoblotting analysis. (d) Knockdown of Npl4 but not Ufd1 stabilizes FNTA. Cells stably expressing FNTA-GFP were transfected with the indicated siRNAs. The level of FNTA-GFP was measured by immunoblotting analysis of immunoprecipitated samples. A fraction of whole-cell extracts were directly analyzed to verify knockdown efficiency.