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. 2022 Feb 9;14(2):344. doi: 10.3390/v14020344

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© 2022 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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Only WT(p27)-Rex interacts with WT-Upf1. (A). Hyperphosphorylated and RNA helicase-deficient Upf1 mutants were generated (upper panel) and their interaction with WT-Rex was examined by GST pulldown assay in GST-Upf1 and FLAG-Rex-expressing HEK293FT cells. The results showed that Rex interacted only with WT-Upf1 and not with the NMD-deficient Upf1 mutants (lower panel). (B). The interaction of WT-Rex and p21Rex with WT-Upf1 was examined by GST pulldown assay in GST-Upf1 and FLAG-Rex-expressing HEK293FT cells. The results showed that only WT-Rex interacted with Upf1 (bottom left), which was correlated with NMD inhibitory capacity measured by the NMD activity reporter assay system in HeLa cells (bottom right) (n = 6, mean ± SD, ** p < 0.01).