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. 2005 Oct 11;1(2):66–78.

Figure 1.

Figure 1.

A siRNA targeting the HIV-1 promoter inhibits HIV-1 replication: (A) Location of sequence targeted by the siRNA. The positioning of NF-κB binding sites in the 5′LTR are indicated by red dots; the blue shading represents the target sequence of the siRNA (HIV-prom-A); the underlining indicates the sequence of the tandem NF-κB binding motifs. (B) Effect of short siRNAs on the time course of HIV-1 (NL4-3) production in MAGIC-5 cells: At day 7 siRNAs were transfected into productively infected cells (indicated by arrow). Reverse transcriptase (RT) levels are shown for cultures transfected with HIV-prom-A siRNA (blue triangle) and scrambled siRNA (red circle), mock transfected cells (green square), and uninfected cells (black square). (C) HIV-1 proviral DNA is present in HIV-prom-A siRNA transfected cells. Provirus was detected by PCR amplification of 154bp fragment of HIV-1 gag gene from DNA extracted from MAGIC-5 cells 25 days after infection. NC indicates non-infected MAGIC-5 cells. (D) Abrogation of HIV-1 protein synthesis induced by HIV-prom-A siRNA shown by western blot analysis of HIV-Gag-p24, Gag precursor protein p55 and beta-actin expression in cell lysates of MAGIC-5 cells 10 days after infection with NL4-3. (E) HIV-1 RNA is markedly reduced in the nuclei of cells transfected with HIV-prom-A siRNA. Viral RNA was quantified by real time PCR in the nuclei isolated from MAGIC-5 cells either 10 days or 20 days after infection. HIV viral RNA copy number is shown as copies per 1000 copies of beta-actin. NC indicates non-infected MAGIC-5 cells.