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. 2001 Oct 23;98(22):12695–12700. doi: 10.1073/pnas.221134198

Figure 4.

Figure 4

(A) Cell-surface CXCR4 and CCR5 expression by uninfected PHA-stimulated PBMC exposed to 0 or 10 μM norepinephrine for 24 h (similar effects were observed in HIV-1-infected PBMC). Vulnerability to infection was assessed by real-time PCR quantification of proviral R/U5 long-terminal repeat structures (relative to human β-globin controls) in total cellular DNA isolated 12 h. after exposure to NL4–3 or Ba-L at 0.05 multiplicity of infection (similar effects observed with SF-162 and Ada-M). (B) Effects of norepinephrine on HIV-1 gene expression as quantified by flow cytometric detection of a virus-encoded murine heat-stable antigen (mHSA/CD24) reporter gene in PHA-stimulated PBMC treated with 0, 1, or 10 μM norepinephrine for 24 h after infection with HIV-1NL-r-HSAS (20). To verify that differences in immunofluorescence reflected increasing reporter gene density on infected cells (rather than increasing numbers of infected cells), anti-mHSA fluorescence intensity was quantified on mHSA+ cells (gating out mHSA− cells falling below maximum isotype control fluorescence).