The antiviral activity of primate Viperin orthologs was assayed. (A) 293T cells were co-transfected with HIVLaiΔNef that encodes a luciferase reporter gene (200 ng) and the indicated primate Viperin (700 ng). Virus yield was measured by infecting SupT1 cells and assayed for luciferase expression. Error bars represent standard deviations of four infection replicates and are representative of three independent experiments. (B) Single-cycle infectious virus yield of VSV-G pseudotyped primate lentiviruses from 293T cells co-transfected with 700 ng of human Viperin, rhesus Viperin or empty vector was titered on TZM.BL indicator cells. The infectivity readout by β-galactosidase activity was measured in relative light units (RLU). Error bars indicate standard deviations of four infection replicates, and are representative of at least three independent experiments.