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Figure 4 | Retrovirology

Figure 4

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

Figure 4

Contribution of SAMHD1 nuclear localization to restriction of HIV-1. Human monocytic U937 cells stably expressing the indicated mutant and wild type SAMHD1 proteins (A) were challenged with increasing amounts of HIV-1-GFP (B). As a control, U937 cells stably transduced with the empty vector LPCX were challenged with HIV-1-GFP. (C) U937 cells stably expressing the indicated SAMHD1-FLAG variants were fixed and immunostained using antibodies against SAMHD1 (green). The cellular nuclei were stained by using DAPI (blue). Mock represents wild type U937 cells. Image quantification for three independent experiments is shown in Table 1. (D) SAMHD1-silenced THP-1 cells were transiently transduced with the indicated wild type and mutant SAMHD1 proteins. Forty-eight hours post-transduction cells were differentiated and infected with HIV-1 LUC-G, which expresses luciferase as a reporter for infection. Luciferase activity was measured 24 hours post-infection. Results are expressed as fold increase luciferase activity in transduced over mock THP-1 cells. (E) The level of mutant and wild type SAMHD1 expression, in transduced SAMHD1-silenced THP-1 cells, was analyzed by Western blotting using anti-FLAG antibodies. As loading control, we analyzed cell extracts by Western blot using anti-tubulin antibodies. Similar results were obtained in three independent experiments and a representative experiment is shown.

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