FIG 7.
Cotransfection of μNS and T3v10M1 μ2 promotes progeny production. (A) H1299 cells were cotransfected with μNS and wild-type (wt) μ2 or μ2 derived from T3v10M1 (10M1). The μ2 were either C- or N-terminally Flag-tagged as indicated. At 24 h posttransfection, cells were infected at an MOI of 3 with T3wt, and whole-cell lysates were harvested at 12 h postinfection. Immunoprecipitation was performed using anti-μNS or anti-Flag, as indicated. Western blot analysis was then conducted with anti-μ2 or anti-μNS (α-μNS). Relative levels of Flag-μ2 coprecipitated with μNS, or reciprocally, μNS coprecipitated with Flag-μ2, are provided below the corresponding lanes. Protein band intensities were quantified by ImageQuant. (B) H1299 cells were transfected with a plasmid expressing either an N- or C-terminal Flag-tagged μ2 derived from T3wt or from T3v10M1 (10M1). After 24 h of transfection, cells were infected with T3wt at an MOI of 4. At 9, 12, and 20 hpi, whole-cell lysates were frozen and thawed three times and subjected to plaque titration. Data are presented as the ratio of infectious virus (PFU) from T3wt-infected cells transfected by 10M1-derived μ2 over T3wt-infected cells transfected with T3wt-derived μ2 (mean ± SD; one-sample t test comparing sample mean to theoretical mean of 1). *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.