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The objective was to determine the differential redifferentiation characteristics of human articular chondrocytes (HACs) in monolayer, alginate beads and pellet culture by investigating mRNA expression, protein expression, GAG content and cell proliferation.
The human immunodeficiency virus (HIV) Rev protein is essential for viral structural protein expression (Gag, Pol, and Env) and, hence, for viral replication.
Additional post-transcriptional defects were observed at the levels of Gag expression, Gag processing, Gag release and virus infectivity.
Although we were able to overcome blocks in the HIV replication cycle in murine cells up to proviral integration, we observed defects in transcription, despite expression of the hCyclin T1 transgene, and after transcription at the levels of Gag expression, Gag processing and Gag release.
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The percentage of NYVAC-C-KC infected cells expressing Gag was also decreased at 24 hours post infection, though there was considerably more Gag expression compared to that in cells infected with NYVAC-C.
As shown in Figure 2B, the treatment with PonA alone resulted in inefficient expression of Gag products in 18-4s cells, while more than 50% of the parental 293/RGP cells was induced to express Gag products.
The ability of NF90ctv and its deletions to bind RRE was determined by Western blot of the expression of Gag, using HIV-1IIIB p55 Gag antibodies (obtained through the NIH AIDS Research and Reference Reagent Program, Division of AIDS, NIAID, NIH).
If HIV-1 protease activity in virus producing cells is responsible for the activation of CASP3*, then the expression of Gag-pol, not Gag, should confer the cell growth disadvantage, resulting in reduced levels of Gag in these cells.
Finally, the cells were harvested and the level of expression of Gag was evaluated by Western blot, using Gag antibodies.
In mouse T cells infected with an efficiency of 20 25%, expression of Gag was reduced approximately 70-fold compared to human T cells (Figure 8A) and Gag remained unprocessed (Figure 7B).
Nullbasic downregulated the expression of Gag and Env from a HIV-1 proviral plasmid that expressed viral mRNA from a CMV promoter (Fig. 3C).
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com