FIG. 5.

trans-complementation of M1 (μ2-specific) RNAi with μ2 proteins containing mutations in a conserved N-terminal polybasic region. (A) Substitutions in a basic amino acid domain. Changes are shown relative to the wt sequence encoded by the parental vector, pM1T1L. (B) Viral growth in cells expressing vector-encoded μ2 proteins. Parental or M1 shRNA-expressing 293T cells were transfected with empty vector (pcDNA) or T1L μ2-expressing vectors, followed by infection with T3D at an MOI of 10 PFU/cell. At 24 h postinfection, viral titers in culture lysates were determined by plaque assay. Results are mean viral titers from three independent experiments. Error bars denote standard deviations. (C) Confirmation of μ2 expression in transfected cells. Expression of μ2 proteins from trans-complementation vectors was verified by immunoblotting of protein extracts from transiently transfected cells using μ2-specific antiserum.