FIG. 6.

Nuclear localization and integration of reverse transcription intermediates in resistant cell lines. (A) Cells (5 × 10⁵/well of a six-well plate) of the indicated cell lines were infected as described for Fig. 5, and nuclear DNA was harvested at 0 hpi and 24 hpi. QPCR was performed using a primer/probe set that recognizes the second-strand transfer reverse transcription intermediate. (B) Cells (5 × 10⁶/6-cm dish) of the indicated cell lines were infected with 5 × 10⁶ IU of LEGFP[VSV-G]. DNA was harvested 24 hpi, and QPCR was performed with a primer/probe set that recognizes 2LTR circles. (C) Cells (5 × 10⁵/well of a six-well plate) of the indicated cell lines were infected with 5 × 10⁵ IU of pLEGFP[VSV-G]and passaged for 10 days postinfection. Total DNA was isolated, and QPCR was performed using a primer/probe set that recognizes the first-strand transfer (+SSS) and subsequent reverse transcription intermediates. DNA concentration was quantitated by A260, and number of DNA molecules was determined by comparison to a standard curve generated from serial dilutions of plasmid DNA. The data shown are the average mean values obtained in an experiment with triplicate samples and are representative of results of three independent experiments. Error bars indicate the standard deviations of the data.