Figure 2
TM domain and GPI domain of human tetherin are important for the inhibition of PFV. (A) Schematic representations of human tethrin (HuTHN) and deletion mutants. WT, wild type; delCT, HuTHN (20-180 aa); delTM, HuTHN (47-180 aa); delGPI, HuTHN (1-161 aa). (B, C) pcPFV was transfected into 293T cells together with the indicated HuTHN wild type or truncated mutants. The amount of virus in supernatants was analyzed by infecting PFVL (B), Western blotting (B), and viral DNA copy number assay (C). The luciferase activity and viral DNA copy number were normalized to the samples transfected with empty vectors (control). The cell lysates were immunoblotted with anti-Gag antibody. The same blots were reprobed with anti- β-actin antibody and anti-FLAG antibody. The viral particles were immunoblotted with anti-Gag antibody and Odyssey goat anti-rabbit IRDye 680. The intensity showed below the blots was performed with LI-COR analysis software.