Figure 6. AA inhibits SV40 DNA replication in normoxically cultivated permeabilized cells.

(A) SV40-infected cells were permeabilized and incubated normoxically in the presence or absence of AA (2 μM). Cultures were labelled for 5 min with [α-³²P]dATP (10 μCi/ml, 0.1 μCi/nmol dATP), 30 min after permeabilization, and processed for alkaline sedimentation on a 15–30% sucrose gradient. Sedimentation was from left to right; the top and bottom of the gradients are indicated. (B) Formation of SV40 form U. SV40-infected cells were permeabilized and incubated normoxically in the presence or absence of AA (2 μM) or hypoxically for 30 min. For the last 15 min, aphidicolin (2 μg/ml) was added. Thereafter, incubation was stopped and isolated viral DNA was separated on a chloroquine-containing agarose gel, blotted and detected using ³²P-labelled SV40-DNA as probe. LC, late Cairns SV40 DNA; IC, intermediate Cairns SV40 DNA; T, topoisomers of mature SV40 DNA (form I); U, form U.