Fig. 1

Mouse SAMHD1 inhibits lentiviral infection at the level of reverse transcription. a Bone marrow derived dendritic cells (BMDC) from wild-type (WT; n = 3) and SAMHD1 knockout mice (KO; n = 3) were either not infected (mock) or infected with VSV-G pseudotyped HIV-CMVGFP reporter virus (MOI of 1). Reporter virus infection was quantified 3 days later by flow cytometry. The percentage of infected cells (GFP+) is shown as average of triplicate infections with error bars indicating the standard deviation. One out of two independent experiments is shown. b PMA-treated U937 cells expressing murine isoform 1 (iso1), isoform 2 (iso2), or empty control vector were infected with VSV-G pseudotyped HIV-E2-Crimson reporter virus (MOI of 1). The percentage of infected cells has been determined 3 days later by flow cytometry. The columns represent the average of three independent experiments with error bars indicating the standard deviation. To control for SAMHD1 iso1 and iso2 expression, cell lysates were prepared and analyzed on an immunoblot probed with anti-HA MAb or anti-GAPDH antibody. c BMDC from WT (n = 3) or KO mice (n = 3) were incubated VSV-G/HIV-CMVGFP reporter virus at a MOI of 1. Cellular DNA was isolated 12 and 24 h postinfection and used as a template in qPCR to amplify HIV-CMVGFP reverse transcription products. Nevirapine (NVP) was added to the indicated wells 14 h prior to infection to control for contaminating plasmid DNA. The data are presented as the average of triplicates with error bars indicating the standard deviation. One out of three independent experiments is shown. d PMA-treated U937-control, U937-iso1, and U937-iso2 cells were incubated VSV-G/HIV-CMVGFP reporter virus at a MOI of 1. Total DNA was isolated from the cells at 12 and 24 h postinfection and used to amplify reverse transcription products by qPCR. The data are presented as the average of triplicates with error bars indicating the standard deviation. The results shown are representative of results obtained in at least three independent experiments