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. 2011 Jun;85(11):5483–5493. doi: 10.1128/JVI.00254-11

Fig. 8.

Fig. 8.

Cav-1 modulates HIV-1 gene expression in physiologically relevant primary monocyte-derived macrophages (MDMs). (A and B) MDMs were isolated from human peripheral blood. Cells were dually infected with VSV-G-pseudotyped NL4-3.Luc.R−E− and Ad-Cav-1 or Ad-GFP. The expression levels of GFP and Cav-1 are shown. (C) The level of viral gene expression in the presence of overexpressed or endogenous Cav-1 was measured by luciferase activity in one round of replication and determined relative to the levels in cells transduced with Ad-GFP. (D) To determine the physiological relevance of the inhibition of HIV gene expression by Cav-1, MDMs infected with VSV-G-pseudotyped NL4-3.Luc.R−E− were transfected with specific siRNA that targets Cav-1 or control (CTL) siRNA using Fugene 6. (E) The level of viral gene expression in one round of replication was measured by luciferase activity under the viral LTR promoter. (F, G, and H) In separate experiments, MDMs were transfected with pLTR-GFP or NF-κBM2 (F), NFAT5M (G), or Sp1M (H) along with pCZ-Cav-1 or pCZ-vector to examine the role of NF-κB in Cav-1-mediated inhibition of HIV gene expression in macrophages. The cells were harvested 48 h posttransfection and subjected to flow cytometry to measure GFP fluorescence intensity. All experiments were performed in triplicates, and results shown are means ± SD with P values. CTL, control.