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Fig. 5 | Molecular Medicine

Fig. 5

From: Nuclear Localization Signal of HIV-1 as a Novel Target for Therapeutic Intervention

Fig. 5

PCR-based analysis of nuclear translocation of HIV-1 genome

(A) Results of the PCR analysis with primers specific for HIV-1 polymerase gene (pol) and 2-LTR circle junction (2-LTR circle) or for the cellular α-tubulin gene. A million monocytes were infected with HIV-1ADA in the presence of H-0294 (2) or H-2194 (21). Total cellular DNA was extracted immediately after a 2-hr adsorption period (0 hr p.i.), 48 hr p.i. and 96 hr p.i. HIV-1-specific products were revealed after Southern hybridization to 32P-labeled probe, while the tubulin-specific fragment was revealed by staining the gel with ethidium-bromide. Control infection (C) was performed without drugs. Results of the PCR reactions with serial dilutions of 8E5 cells, each containing one integrated copy of HIV-1 genome (53), using pol-specific primers are shown on the right. The number of HIV-1 copies in each dilution is shown above the corresponding lane. (B) Results of the experiment described in Fig. 4A were analyzed on a phosphorimager. Nuclear translocation index is expressed as \(\left( {\begin{array}{*{20}c} {{{{{{{\rm{N}}_{2 - {\rm{LTR}}}}} \over {{{\rm{N}}_{{\rm{tot}}}}}}} \over {{{{{\rm{C}}_{2 - {\rm{LTR}}}}} \over {{{\rm{C}}_{{\rm{tot}}}}}}}}} \\ {} \\ \end{array} } \right)\begin{array}{*{20}c} { \times 100\% .} \\ {} \\ \end{array} \)

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