FIG. 4.
Analysis of reverse transcription in monocytes (day 1) or mature macrophages (day 9). Cells were harvested at 2-h intervals after exposure to CMVΔR8.9(VSV-G) packaged vector at an MOI of 10. PCR was then performed for intermediate reverse transcripts containing eGFP sequences (A) and second-strand-transfer products containing LTR/gag sequences (B), and serial twofold dilutions were used for PCR of cellular β-actin for a loading control (C). PCR was also performed on supernatants containing an equivalent amount of virus to check for DNA contamination in the viral preparations (lanes V). Serial twofold dilutions of samples from the the 8-h time points of days 1 and 9 were used for PCR to estimate relative amounts of eGFP and LTR/gag transcripts (D). The reverse transcriptase inhibitor zidovudine (AZT) was added to day 1 cells exposed to CMVΔR8.9(VSV-G) packaged vector at an MOI of 10. After 14 h, PCR was performed on cell lysates to detect eGFP transcripts (E). Un, uninfected cells; PI, postinfection.