Fig. 2

Murine SAMHD1 blocks reverse transcription of MLV in a myeloid cell line and primary BMDCs. a BMDC were incubated with VSV-G-pseudotyped MLV-GFP reporter virus with (MLV +PBS) or without (MLV −PBS) an intact Primer Binding Site (PBS) to control for contaminating plasmid DNA (MOI of 1). At 12 and 24 h postinfection, cellular DNA was harvested and MLV reverse transcripts were quantified by qPCR using oligonucleotides targeting the reporter gene sequence. The data are presented as the average of triplicates with error bars indicating the standard deviation. The data for one of three independent experiments is shown. b PMA-treated U937-control, U937-isoform 1, and U937-isoform 2 cells were incubated VSV-G/HIV-CMVGFP reporter virus at a MOI of 1. Cellular DNA was isolated at 12 and 24 h postinfection and used to amplify reverse transcription products by qPCR. The oligos used in qPCR were targeting the reporter gene sequence. 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 three independent experiments. c BMDC from WT (n = 3) and SAMHD1 KO (n = 3) mice were either not infected (mock), infected with VSV-G/MLV-GFP, or infected with VSV-G/HIV-CMVGFP reporter virus at a MOI of 1. Viral infectivity was quantified 3 days postinfection by flow cytometry. The percentage of GFP + BMDC is shown as average of triplicate infections. One out of two independent experiments is shown