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. 2017 Apr 19;7:966. doi: 10.1038/s41598-017-01097-5

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© The Author(s) 2017

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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Hetero-dimerization of E proteins from different flaviviruses. (a) Schemes of sE-cvD constructs used. For all DENV serotypes and ZIKV, sE-cvD was SV5-tagged, while 3sE-cvD was also tagged with BAP and co-expressed with the SV5-tagged proteins. Expected homo- and hetero-dimers and their relative migration in non-reducing PAGE are indicated. (b) Non-reducing western blot of cell extracts (E) and culture supernatants (S) of HEK-293T cells co-transfected with the indicated constructs and developed with anti-SV5 (upper panel) or StrepAv-HRP (lower panel). *Indicates biotinylated host intracellular proteins. Arrowheads indicate migration of dimers as shown in (A). Open arrowheads indicate hetero-dimers. Right panel, long exposure of the poorly expressed 1sE-cvD-SV5 co-transfected with 3sE-cvD-BAP. (c,d) Cytofluorimetry of HEK-293T cells co-transfected with the membrane displayed (m)sE-cvD proteins (c) or (m)sE-wt proteins (d) of all four DENV serotypes or ZIKV and the secretory SV5-tagged 3sE-cvD (c) or 3sE-wt (d). Profiles correspond to anti-SV5 reactivity and are compared to HEK-293T cells transfected only with the secretory SV5-tagged 3sE.