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Figure 4 | BMC Evolutionary Biology

Figure 4

From: Synonymous site conservation in the HIV-1 genome

Figure 4

Gag expression and virion release of the NLR+GFP and NLR+GFP polmutclones in transfected and infected cells. (A) HEK293T cells were transfected with plasmids expressing the VSV-G envelope, and NLR+GFP (WT) or two identical clones of NLR+GFPpolmut (mut1, mut2). Mock represents control cells transfected with no plasmid DNA. Gag precursor (Pr55gag) was detected in extracts of transfected cells (two days post transfection) by Western blotting using anti-capsid monoclonal antibody. Actin was used to control for protein levels in the samples. (B) Virions were purified from equal volumes of supernatants of cells in (A) and their levels were determined by detecting the capsid protein (p24), using Western blotting as above. (C) Equal amounts of virions from (B), normalized by RT activity, were used to infect naïve 293T cells and the newly infected cells were analyzed two days post infection as in (A). (D) Virions from supernatants of the cells indicated in (C) were analyzed as in (B). For (A-D), one representative Western blot is shown at the top of each panel and bars (bottom part) represent the average densitometry of the bands from three independent experiments.

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