Fig. 2
UPF2 and SMG6 knockdown enhance HIV-1 viral gene expression and replication in primary MDMs. Human monocytes were differentiated into MDMs and then transfected with the indicated siRNAs. After 24 h, cells were infected with NL4-3-Bal-IRES-HSA virus (MOI: 1.0) and kept in culture for 6 days. Cells silenced for a UPF1, b UPF2 or c SMG6 were collected, lysates were run on SDS-PAGE gels and protein levels were detected by Western blotting. d 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. Asterisks represent statistically significant difference between groups (One-way ANOVA; ns: not significant). e Cells silenced for UPF1, UPF2, SMG6 or SAMHD1 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 5 different donors each. Asterisks represent statistically significant difference between groups (One-way ANOVA; ns: not significant, *p ≤ 0.05 and p ****p ≤ 0.0001). f Representative dot plot depicting HSA expression in siNS, siUPF1, siUPF2, siSMG6 and siSAMHD1 transfected primary MDMs