Fig. 6.

Mouse SAMHD1 does not affect viral RNA level in infected murine cells. a PMA-treated U937-control, human SAMHD1 (huSAM), mouse isoform 1 (iso1), and mouse isoform 2 (iso2) expressing cells were incubated with VSV-G/HIV-E2-Crimson reporter virus at a MOI of 1. At several time points postinfection (hours post infection, hpi), the infected cells were lysed and total cellular RNA was isolated, quantified and reverse transcribed. HIV transcripts within the cDNA were quantified by qPCR using oligos targeting the E2-Crimson reporter gene. PCRs on cDNA samples generated in the absence of a reverse transcriptase were used to control for plasmid contamination (not shown). The amounts of HIV transcripts are shown as percent of the HIV RNA content at 1 h postinfection. The average of triplicate infections is shown with error bars indicating the standard deviation. One out of three independent experiments is shown. b BMDC from SAMHD1 KO (n = 3) or WT (n = 3) mice were infected with VSV-G/HIV-E2-Crimson reporter virus at a MOI of 1. Similar to (a), viral RNA from uninfected cells (−) or infected cells was isolated at various time points postinfection, quantified and is depicted as number of HIV RNA copies/100 ng input RNA. The columns represent the average of triplicate infections with error bars indicating the standard deviation. One of two independent experiments is shown. c The average amount of viral RNA copies in infected cells from WT (n = 3) or KO mice (n = 3) at different time points postinfection as shown in (b). HIV transcripts were quantified relative to the maximum amount of transcripts at 3 h postinfection