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Figure 3 | Retrovirology

Figure 3

From: Triptolide inhibits human immunodeficiency virus type 1 replication by promoting proteasomal degradation of Tat protein

Figure 3

Triptolide inhibits HIV-1 transcription. (A) Time-of-addition analysis. TZM-bl cells were infected with HIV-1 (MOI = 0.5, strain NL4-3), and the virus inoculum was removed after 1 h of absorption. Test compounds (dextran sulfate at 10 μg/mL, zidovudine at 1 μM and triptolide at 5 nM) were added at the indicated time points after infection. Reporter signal was monitored at 48 h post-infection as a measure of viral replication. (B) The effect of triptolide on HIV-1 proviral DNA formation. Jurkat cells were infected with HIV-1 (MOI = 0.5, strain NL4-3) in the presence of the test compounds (INI 118-D-24, 50 μM; flavopiridol, 20 nM; and triptolide, 5 nM). Proviral DNA synthesis was determined by nested PCR analysis at 12 h post-infection. (C) The effect of triptolide on HIV-1 mRNA synthesis. Jurkat cells were infected with HIV-1(MOI = 0.5, strain NL4-3) in the presence of the test compounds (INI 118-D-24, 50 μM; flavopiridol, 20 nM; triptolide, 5 nM). Viral mRNA (the Gag region) content was determined by reverse-transcription PCR analysis 12 h post-infection. (D) Inhibition of luciferase expression encoded in pNL4-3.Luc.R-E- by tiptolide. HIV-1 molecular clone pNL4-3.Luc.R-E- and pRL-TK (transfection efficiency control) were co-transfected into Jurkat cells. Transfected cells were cultured in the presence of the test compounds (INI 118-D-24, 50 μM; flavopiridol, 20 nM; triptolide, 0.2 ~ 5 nM) for an additional 24 h before measuring luciferase activity with a dual-luciferase assay system. Results were presented as the mean plus the standard deviations (n = 3).

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