Fig. 1

SAMHD1 and miRNAs in vitro and in vivo. a miRNAs suppress SAMHD1-linked reporter assay. 293T cells in 96-well plates were transfected with 10 nM scrambled RNA control (scRNA) or 10 nM miRNA mimics (Life Technologies), along with 50 ng luciferase reporter vector containing the human SAMHD1 3′ UTR (SwitchGear Genomics) or empty vector. After 48–72 h, luciferase activity was measured using a Fluoroskan Ascent fluorometer (Thermo Scientific). b–d Normalized fold change of thalamic SAMHD1 and miR-181a expression compared with the average of uninfected controls. Total RNA was extracted from the archived thalamus samples from an SIV model of HIV disease. SAMHD1 levels were normalized to the average of GAPDH and actin beta expression using data collected using hydrolysis probe qPCR assays (Life Technologies). Expression of miR-181a (shown) and other miRNAs (including miR-34a and miR-155, not shown) were determined by stem-loop reverse transcription/qPCR assays (Life Technologies) and normalized to U6 snRNA. Correlations of miR-181a and SAMHD1 expression are shown for b subjects during acute phase infection (4–14 days post infection), c subjects during progression to disease (21–84 days p.i.), and d for all samples, including uninfected controls. Each point represents one subject. No correlations were significant (p < 0.05) or trended towards significance (p < 0.1)