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Figure 2 | Retrovirology

Figure 2

From: Caveolin-1 reduces HIV-1 infectivity by restoration of HIV Nef mediated impairment of cholesterol efflux by apoA-I

Figure 2

Cav-1 restores HIV −1 Nef mediated impairment of cholesterol efflux to apoA-I. U87-CD4-CXCR4 cells were transfected with Nef expression plasmid (pcDNA3.1SF2Nef) along with pCZ-cav-1 or pCZ vector. Cells were labeled with [3H] cholesterol for 36 h. (A) The level of Nef and Cav-1 expression was determined by Western blot analysis. (B) ApoA-I induced cholesterol efflux is shown. All experiments were performed in triplicate, and results shown are mean ± SD with P values. (C) Primary monocyte derived macrophages (MDMs) were infected with Ad-Cav-1 or Ad-GFP, followed by infection with VSV-G pseudotyped HIV, either carrying Nef (psHIVwtNef) or defective Nef (psHIVΔNef) at an moi of 5. ApoA-I mediated cholesterol efflux was performed and analyzed by incubating cells in medium in the presence or absence of 50 μg/ml apoA-I. The results are presented as a percentage of cholesterol efflux to apoA-I from control (set as 100%), and are the mean± SD of triplicate determinations. The expression levels of Cav-1 and GFP are shown in (D). (E) Primary macrophages were infected with AD8 or nef defective AD8 (ADnefmut) at an moi of 0.01. ApoA-I mediated cholesterol efflux was measured 15 days post infection. (F) THP-1 cell-differentiated macrophages were infected with psHIVwtNef or psHIVΔNef at an moi 3 with or without Ad-Cav-1. On day 5 after infection, Oil red O staining was performed and a representative area in each well is shown.

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