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. 2020 Oct 22;10:18101. doi: 10.1038/s41598-020-75118-1

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© The Author(s) 2020

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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NFC enables rapid detection of mutations affecting PR processing efficiency and virus release. (A) PR processing of VIPER with wild-type NL4-3, IN D116A, IN E152A, IN H12N, and PR D25N mutants, analyzed using NFC. The IN H12N and PR D25N mutations both significantly reduced processing efficiency (p < 0.0001 for both). (B) Western blot analysis of p24 CA processing with wild-type NL4-3 and mutant viruses. The PR D25N mutant resulted in severe disruption of Gag and Gag-Pol processing as expected for the catalytic site mutation. The IN H12N and E152A mutants resulted in slight elevations in unprocessed Gag and processing intermediates. (C) Release of HIV-1 viruses as measured by NFC. The IN E152A and H12N mutations resulted in significant reductions in virus release (p < 0.0001 for both). The PR D25N mutation resulted in a modest but significant increase in virus release (p = 0.021). (D) Virus release for the E152A mutant was fully restored by treating cells with saquinavir. However, the H12N mutant continued to demonstrate significantly reduced processing (p < 0.001) in the presence of saquinavir. Virus release in the absence of saquinavir is replotted from the data shown in part (C).