Figure 6.

Quantification of LTR circles, linear unintegrated, and integrated HIV-1 DNA by Southern blotting and Alu-PCR confirms defects in nuclear import for all DNA Flap mutants. (A) MT4 cells were infected with wild-type or DNA Flap mutant viruses pseudotyped with VSV-G envelope. Total DNA was isolated 48 h post-infection and intracellular viral DNA forms were analysed by Southern blotting. Bands were quantified using Phosphorimager and nuclear import was assessed by plotting the ratio of LTR circles (nuclear) to linear non-integrated HIV-1 (predominantly cytoplasmic). (B) Assessment of HIV-1 integration by Alu-PCR. The graph shows mean values of 3 independent experiments +/- SEM.