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. 2020 Sep 16;12(9):1032. doi: 10.3390/v12091032

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© 2020 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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Delayed transient recruitment of ALIX into HIV Gag (YP⁻ + PTAP⁻) VLPs or HIV Gag (PTAP⁻) VLPs. HeLa cells stably expressing ALIX–h30–eGFP were transfected with 1500 ng of Gag (PTAP⁻)–mCherry (A,B) or Gag (PTAP⁻ + YP⁻)–mCherry (C,D) and imaged 5 h after transfection. Intensity plots (as gray dots and fitted with gray line) of individual fully assembled representative VLPs are shown and cropped TIRF images of the Gag (top, gray) and AIX (bottom, blue) for (A) Gag(PTAP⁻)–mCherry VLPs and (C) Gag (PTAP⁻ + YP⁻)–mCherry VLPs. Histograms of the first time (dark blue) and later recruitments (light blue) of ALIX are shown for (B) Gag (PTAP⁻)–mCherry VLPs and (C) Gag (PTAP⁻ + YP⁻)–mCherry VLPs. Alix was recruited within 2 h after the start of the actual experiment.