Fig. 3
From: HIV-1 Penetrates Coronary Artery Endothelial Cells by Transcytosis
HIV-1 R/U5 reverse transcription in CAEC. Two hundred fifty thousand CAEC in a 6-well plate were infected with HIV-1LAI at the multiplicity of 0.2, 4, or 40 RNA copies per cell and HIV-1JR-FL at 40 copies per cell. The virus stocks were pretreated with 400 U/mL RNase-free DNase I, as described in Materials and Methods. PCR lysates were prepared 7 days PI and were subjected to PCR amplification using primers from the LTR/U5 region, the pol gene, or 2 LTR, and Southern blot hybridization with the appropriate P32-labeled probe. Serial dilutions of 8E5/LAI cells, containing one provirus per cell, were amplified with the same set of LTR/U5 primers as PCR standards. Reverse transcripts were detected at 7 days PI using only the LTR/U5 primers (data shown). No hybridization was obtained using the pol and 2LTR primers (data not shown).