Figure 2

Effects of dominant-negative mutants of Nedd4-like E3 ubiquitin ligases on RD-114 production. (A) 293 T cells were cotransfected with pTERD-114 (100 ng) and either the expression plasmid for the dominant-negative mutants of Nedd4-like E3 ubiquitin ligases, which express only WW domains containing N-terminal FLAG-tag, or the empty vector pCDNFL (Control) (1 μg). Cells and viruses were collected at 72 h after transfection, and analyzed by Western blotting (upper) and real-time RT-PCR (lower). The virus production in Control was set to 100%. The data are shown as averages and standard deviations of 3 independent experiments. (B) CRFK cells were transfected with the expression plasmid for the dominant-negative mutant of Nedd4-like E3 ubiquitin ligases (1 μg) and analyzed as in (A). Statistical analysis was performed by using the Student's t-test (*: p < 0.01)