Figure 2From: Vpx rescue of HIV-1 from the antiviral state in mature dendritic cells is independent of the intracellular deoxynucleotide concentrationVpx degrades SAMHD1 in the presence of the antiviral state but does not increase nucleotide levels. MDDCs were stimulated with 100 ng/ml LPS for 18 hrs and treated with SIVMAC VLPs containing Vpx, VpxQ76A or no Vpx (ΔVpx) for 24 hrs. 0.8 x 105 cells were harvested for SAMHD1 western blot (A). SAMHD1 protein levels were measured and normalized to the actin signal. The ΔVpx sample was set to 100% (B). 2 x 106 were harvested for nucleotide extraction and measurement of intracellular deoxyadenosine concentration (C). 0.5 x 106 were challenged with NL4-3 GFP after 2 hrs of VLP treatment in the presence of LPS (D) or absence (E) and 0.5 x 105 were assessed for GFP expression by flow cytometry 3 days later. 1.6 x 106 cells were treated with either 1 μM MLN4924 or DMSO for 2 hrs prior to Vpx-VLPs or ΔVpx-VLPs addition. 24 hrs later cells were harvested for SAMHD1 western blot (F) or deoxyadenosine concentration measurement for one sample per condition (G).Back to article page