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. 2018 Dec 12;10(12):710. doi: 10.3390/v10120710

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© 2018 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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Influence of the mutation L1P on virus release and Gag-processing. HeLa SS6 cells were transfected with pNLenv1 wt or pNLenv1 L1P, respectively. Cell lysates and VLP fractions were analyzed by Western blotting using a p24-reactive (A) or a p6-reactive (B) antiserum. (C) The efficiency of virus release was calculated as the ratio of Gag (Pr55 and p24) present in the VLP fraction relative to the total amount of Gag detected in cells and released VLPs. (D) The rate of p24 processing was determined by calculating the ratio of p24 vs. Gag (Pr55 and p24) detected in released VLPs. (E) The rate of p6 processing was determined by calculating the ratio of p6 vs. Gag (Pr55, NC-p6, and p6) detected in released VLPs. (C–E) Band intensities were densitometrically quantified with AIDA. Values of pNLenv1 wt were set to 100%. Scattered blots with columns representing mean values of four (C,D) or three (E) independent experiments ± SD. One sample t-test was conducted to determine statistically significant differences in virus release (C), p24 processing (D) and p6 processing (E) between the mutant pNLenv1 L1P and pNLenv1 wt (*p < 0.05; not significant (n.s.) p ≥ 0.05).