Figure 4. Redoxal attenuates virus production and infectivity.
A. Redoxal increases A3G protein levels in HIV-1 producer cells. 293T cells were co-transfected with 0.1 μg A3G-3xHA and 1 μg pNL4-3GFPΔEnv HIV-1 plasmids using VSV-G envelope glycoprotein for pseudotyping. Five hours post-transfection, the media was replaced with fresh media supplemented with DMSO or 1250 nM redoxal. At 40 h post transfection, supernatants containing virus were collected and producer cells were lysed. A3G, Vif, p24Gag, GFP and β-tubulin protein levels in producer cell lysates were analyzed by Western blotting B. Redoxal decreases viral particles produced independently of A3G expression. Virus production from DMSO- and redoxal-treated cells was quantified by RT assay. Percentage of virus production is relative to virus production of untreated producer cells. Results representative of two independent experiments each performed in duplicate. C. Redoxal attenuates infectivity of viruses produced from cells expressing A3G proteins. TZM-bl reporter cells were infected with viruses corresponding to 4,000 RT units. Infectivity was measured 48 h post infection. Results are representative of two independent experiments each done in triplicate. The percentage decrease in the infectivity of viruses produced from redoxal-treated cells compared to controls is shown below the graph. Shown in B and C are means ± SD of duplicate samples from independent wells (p-values from student’s t-test).