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. 2020 May 22;8(2):243. doi: 10.3390/vaccines8020243

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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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Analysis of stability of GC-AG Nef^mut/E7. Comparative stability analysis between Nef^mut/E7 and GC-AG Nef^mut/E7 fusion proteins in total lysates from HEK-293T cells transfected with the respective DNA vectors. Forty-eight hours after transfection, cells were washed and reseeded in complete medium containing 1 μg/mL of cycloheximide. Transfected cells were harvested at the indicated times, and total lysates from the same number of cells were analyzed by Western blot assay. Filters were revealed by both anti-Nef and anti- β-actin polyclonal antibodies. Time zero refers to cells harvested at the time of cycloheximide treatment. Signal intensity ratios compared to respectve time zero are indicated. Molecular markers are given in kDa. The results are representative of two independent experiments. Uncropped blots showing all the bands with all molecular weight markers are shown in Supplementary Figure S2.