Fig. 5

Restriction activity and expression levels of selected Fv1 alleles. a Quantitative western blot analysis of various Fv1 alleles in MDTF-R18 cells transduced with either TYIx-Fv1 or LxIY-Fv1 vectors. Fv1 was detected using an anti-Fv1 antibody and the LI-COR Odyssey infrared fluorescent western blot system. Transduced cells were treated with 0, 10, 100, or 1000 ng/mL doxycycline for 24 h before challenging with EGFP-expressing tester viruses. Quantities of Fv1 in lysate samples were interpolated from a standard curve generated using fluorescent signals from known quantities of purified Fv1NTD protein loaded on the same gel. Numbers under the blot indicate the quantities of Fv1 in each sample relative to that in the LxIY-Fv1^b sample. b Graph showing the mean and standard deviation values of relative quantities of selected Fv1 alleles in MDTF-R18 cells transduced with TYIx-Fv1 or LxIY-Fv1 vector relative to that in the LxIY-Fv1^b sample. Two repeated quantitative western blots were carried for each of two samples of transduced cells, and the results are also listed in Table S2. Cells transduced with TYIx-Fv1 vectors were treated with 0 (black), 10 (red), 100 (blue) or 1000 (green) ng/mL dox for 24 h. Cells transduced with LxIY-Fv1 vectors (purple) were not treated with dox. c–g Scatter plots showing mean values and standard deviation of c N-MLV, d B-MLV, e NB-MLV, f EIAV and g FFV restriction activities from Table 2 against mean relative Fv1 protein levels from Fig. 3b. Blue squares, Fv1^b; red upright triangles, Fv1MIN2; green inverted triangles, Fv1SPR1; purple diamonds, Fv1MAC; orange circles, Fv1CAR1. The two dotted horizontal lines indicate the boundaries between full restriction and partial restriction (0.7), and between partial restriction and no restriction (0.3)