MFI after transfection of U937 clone 10 with antisense oligonucleotides targeting HLE or negative-control β-globin. MFI was determined by flow cytometry using monoclonal anti-CD4-PerCP, monoclonal anti-CXCR4-PE, and polyclonal anti-HLE detected by FITC-conjugated reagents as described in “Materials and methods.” MFI was determined in 2 separate experiments at 36 hours, 48 hours, and 66 hours following antisense transfection to determine minimum reduction in HLE expression. Data represent receptor expression at 48 hours corresponding to the simultaneous HIV-1 infection depicted in Figure 4.

HIV-1 minus-strand, strong-stop DNA in the same cells was detected by digitalization of images following polyacrylamide gel electrophoresis. Variation in PCR amplification was normalized using negative-control β-actin and was calculated as HIV-1 pixels/β-actin pixels. HIV-1 DNA is represented as normalized pixels after HLE antisense/normalized pixels after β-globin antisense. Antisense transfection was performed 3 times and data represent one experiment corresponding to the simultaneous measurement of receptor density.