Not only human tetherin, but also tetherins from simian, bovine and canine inhibit the release of PFV. 293T cells were transfected with pcPFV- along with human tetherin (HuTHN), African green monkey tetherin (AgmTHN), bovine tetherin (BovTHN), or canine tetherin (CanTHN). At 44-48h after transfection, the amount of virus in cell lysates and supernatants was analyzed by infecting PFVL (A), Western blotting (B), and viral DNA copy number assay (C). The FLAG-tag tetherin and Gag proteins were immunoblotted with the antibodies indicated on the left side of each blot. The blot for β-actin was performed as an internal control. The luciferase activity and viral DNA copy number were normalized to the samples transfected with empty vectors (control).