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Figure 5 | Retrovirology

Figure 5

From: Role of SAMHD1 nuclear localization in restriction of HIV-1 and SIVmac

Figure 5

Vpx n -induced degradation of SAMHD1 requires nuclear localization of SAMHD1. (A) HeLa cells were cotransfected with plasmids allowing expression of SAMHD1-FLAG or SAMHD1-K11A-FLAG and HA-tagged Vpx from SIVmac 251(Vpxmac251), SIVrcm-ng (Vpxrcm-ng), HIV-2ROD (VpxROD) or HIV-2B (Vpx2B). Thirty-six hours post-transfection the cells were harvested, and the expression levels of SAMHD1 and Vpx were analyzed by Western blot using anti-FLAG and HA antibodies. As a loading control, cell extracts were Western blotted using antibodies against Tubulin. Similar results were obtained in three independent experiments and a representative experiment is shown. (B) Human monocytic U937 cells stably expressing the indicated mutant or wild type SAMHD1 proteins were challenged with SIVmac or SIVmacΔVpx -GFP reporter viruses. Infection was determined by measuring the percentage of GFP-positive cells. Fold of restriction was calculated by dividing the % of GFP-positive cells resulting from infecting U937 cells containing the empty vector pLPCX by the % GFP-positive cells resulting from infecting U937 stably expressing the indicated SAMHD1 variant. Similar results were obtained in three independent experiments and standard deviation is shown.

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