Figure 2. Murine SAMHD1 Restricts Retro-viral Replication.

(A) BMDCs were generated from six mutant and six control mice and infected with EGFP-containing HIV-1-VSV-G (VSVG/NL43-CMVGFP). Cellular EGFP fluorescence, indicating that the virus had successfully reverse transcribed and integrated its genome, was detected by FACS. The graph represents a summary of two independent experiments with similar results. A third experiment with cells from three additional mutant and three additional control mice yielded similar results (Figure S2). Means ± SD are displayed.

(B) BMDCs from three SAMHD1-deficient mice were transduced with a lentiviral vector carrying either an EYFP and a murine SAMHD1 expression cassette or only the EYFP cassette. After 48 hr, the transduced cells were challenged with the EGFP-expressing HIV-1 pseudotyped with VSV-G (see A). EYFP⁺ cells were analyzed by FACS for EGFP expression 48 hr after challenge. Means ± SD are displayed.

(C) Five SAMHD1^Δ/Δ and five SAMHD1^WT/WT mice were injected intravenously with 1.8 × 10⁷ particles of an EGFP-expressing HIV-1 pseudotyped with VSV-G (VSVG/HR.CMVGFP) and two control mice (Ctrl, one SAMHD1^Δ/Δ and one SAMHD1^WT/WT) were injected with PBS. After 3 days, the mice were sacrificed and the absolute numbers of live EGFP⁺ cells per 10⁶ splenocytes were determined by flow cytometry. Means ± SD are displayed. Differential analysis of individual cell populations from this experiment for HIV reporter infection is shown in Figure S2.

(D) Infection of five additional SAMHD1^Δ/Δ and five additional WT mice with 5 × 10⁶ particles of HIV reporter virus as in (C). Absolute numbers of live EGFP⁺ cells in different splenic cell populations are shown. Means ± SD are displayed.

See also Figure S2.