Figure 6.
Effect of incorporation of gp51 into the virion by mutant forms of the infectious molecular clone pBLV-IF2. COS-1 cells (5.0 × 105) were seeded in a 60 mm dish the day prior to transfection and transfected with 7.6 µg of either wild-type pBLV-IF2, mutant pBLV-IF2, or the control Bluescript II SK (−) vector together with 0.4 µg of pEGFP-N1 using 32 μL of FuGENE HD. (A) Virus particles were collected from the supernatants of the cells and subjected to western blotting analysis with anti-BLV gp51 MAb and anti-BLV p24 MAb followed by horseradish peroxidase-conjugated goat anti-mouse IgG. Positions of the molecular mass marker and the BLV structural protein are indicated. (B) For quantification, densities of bands were analyzed using ImageJ software. Densities of gp51 were normalized with those of p24. Each column and error bar represents the mean ± SD of density for six independent experiments. All values were analyzed by two-way ANOVA with Dunnett’s test. The asterisk indicates a statistically significant difference (* p < 0.05; ** p < 0.01).