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Fig. 5 | Retrovirology

Fig. 5

From: Host mRNA decay proteins influence HIV-1 replication and viral gene expression in primary monocyte-derived macrophages

Fig. 5

Staufen1 knockdown impairs HIV-1 viral gene expression and replication in primary HIV-1-infected MDMs. Human monocytes were differentiated into MDMs and then transfected with control siRNA (siNS) or siRNA against Staufen1. After 24 h, cells were infected with NL4-3-Bal-IRES-HSA virus (MOI: 1.0) and kept in culture for 6 days. a Cells were collected, lysates were run on SDS-PAGE gels and protein levels were detected by Western Blotting. b Fold change in the levels of pr55Gag normalized to the siNS condition. Error bars represent the standard deviation from three independent experiments with cells from three different donors each (One-way ANOVA; p ***p ≤ 0.001). c Cells were collected, incubated with anti-HSA antibody and analysed by flow cytometry. Fold change in the HSA expression was normalized to the siNS condition. Error bars represent the standard deviation from three independent experiments with cells from three different donors each. d Representative dot plot depicting HSA expression in siNS and siStaufen1 transfected primary MDMs. e vRNA was measured by RT-PCR and fold change in the levels of vRNA were normalized to the siNS condition. Error bars represent the standard deviation from three independent experiments with cells from three different donors each (One-way ANOVA; ns: not significant). f Cells were collected at 6 days post-infection, incubated with anti-HSA antibody and sorted through magnetic separation as described in Fig. 1a. Cell lysates were run on SDS-PAGE gels and Staufen1 and actin protein levels were detected by Western Blotting and the fold changes in expression levels between bystander (HSA−) and HIV-1-infected cells (HSA+) were quantified. Error bars represent the standard deviation from two independent experiments with cells from two different donors each. (Student’s t test; ns not significant)

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