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. 2019 Dec 25;12(1):27. doi: 10.3390/v12010027

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© 2019 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 (http://creativecommons.org/licenses/by/4.0/).

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Effect of uncoupled sequential processing on viral infectivity. (a) Representative cytofluorimetry (EGFP) of VERO cells infected with the pseudoviruses produced with the indicated C-Ca-PrME packaging constructs. (b) Quantification of data shown in (a), (N = 5). (c) Western blot of particles produced with the indicated constructs and recovered by ultra centrifugation, and analysed for content of protein E (mAb 4G2) and C (mAb 6F3). (d) Relative quantification of particles recovered with constructs 6 and 7. (e) Western blot of cellular extracts of cells co-transfected with the indicated packaging constructs and without (upper panel) or with (bottom panel) WNV replicon and extracted with sodium carbonate. Supernatants (S, soluble fraction) and pellets (P, membrane associated fraction) were analysed with mAb 6F3 to reveal processed and unprocessed isoforms. (f) Western blot of particles obtained with the indicated constructs and extracted with sodium carbonate (upper panel) or with control PBS (bottom panel). The distribution of the two proteins E and C, in the two fractions (S, soluble and P, membrane associated) were analysed with mAbs 4G2 and 6F3, respectively.