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Figure 4 | Retrovirology

Figure 4

From: Balanced splicing at the Tat-specific HIV-1 3′ss A3 is critical for HIV-1 replication

Figure 4

LNA-mediated masking of ESE tat mimics the influence of the mutated ESE tat on the viral splicing pattern and viral particle production. (A) Schematic illustration of the location of ESS2p, ESEtat, ESE2 and ESS2 as well as the binding site for the locked nucleic acids (LNAs) directed against ESEtat and ESE2/ESS2 sequences. (B) RT-PCR analysis of viral mRNA classes. HeLa cells were transiently transfected with pNL4-3 and either ESEtat-LNA, the ESE2/ESS2 LNA or a scrambled LNA. Total RNA was isolated 24 h post transfection and subjected to RT-PCR analysis using different primer pairs (Figure 1A). (C) Northern blot analysis of total RNA collected in (B) using a DIG-labelled probe hybridizing to HIV-1 exon 7. (D) Western blot analysis of cellular and supernatant viral Gag of co-transfected cells from (B). The detection of actin served as loading control.

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