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. 2019 Mar 26;9:5139. doi: 10.1038/s41598-019-41582-7

Figure 4.

Figure 4

The HIV-1 Rev N-terminus is not required for function but regulates Rev stability. (A) Reporter assay probing export activity of Rev N-terminal truncations. Data are mean ± standard deviation (s.d) of biological replicates. Western blots below show protein expression for Strep-tagged Rev and GAPDH as loading control. (B) Viral replication profiles of viruses containing Rev with N-terminal truncations. Data are mean ± s.d of biological triplicates. Viral p24 levels below 1000 pg/ml are shown as 1000 pg/ml in the plots for illustration purposes. (C) Adding MG132, a proteasome inhibitor partially restores expression of N-terminally truncated Rev in transiently transfected 293 T cells. Numbers above blots represent quantified band intensities for Rev mutants relative to WT-Rev. (D) Shortening the length of N-terminal truncations restores protein expression transiently transfected 293 T cells. Numbers above blots represent quantified band intensities for Rev mutants relative to WT-Rev. (E) N-terminal truncations affect Rev expression/stability from HIV-1 (with Flag-tagged Rev in the nef locus) in infected T-cells. Plot shows virus production from cell supernatant, quantified by p24 ELISA. Western blots below show protein expression for Flag-tagged Rev and GAPDH loading control from the cells. The original blot for the cropped bands is shown in Fig. S5. Data are mean ± s.d of biological replicates.