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. 2020 Nov 10;10:19416. doi: 10.1038/s41598-020-76375-w

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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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Effects of patient-derived Nef clones on SERINC5 downregulation and counteraction of SERINC3/5-mediated infectivity inhibition. (A) Representative flow cytometry plots display the expression of surface SERINC5 in JTAg-SERINC3/5^−/− cells that had been co-transfected with genes encoding GFP and Nef _SF2-GFP together with SERINC5-iHA or an empty vector. The resultant cells were analyzed for cell-surface SERINC5 expression (as HA) and GFP. MFI values for SERINC5-iHA in the GFP⁺ subset are shown. (B) Nef clones isolated from the indicated HIV-infected subjects (2 or 3 clones per patient) were analyzed for their ability to downregulate SERINC5-iHA. Results are expressed as relative downregulation activity, normalized to the control strain, Nef_SF2. Plots indicated in red were used for functional analyses in panel C and Fig. 5. (C) Correlation analysis between Nef’s ability to counteract SERINC3/5 (for viral infectivity) and to downregulate cell surface expression of SERINC5. Nef clones tested are indicated in (B). Statistical analysis was done with the Spearman test.