FIG 7.
Enhancing effect of L305A and dominant negative effect of L302A on wild-type M VLP release. (A) Association of wild-type M with mutants L302A and L305A. 293T cells were transfected with the indicated plasmids. At 48 h posttransfection, the cells were harvested and subjected to coimmunoprecipitation assay as described in the legend to Fig. 3. (B) Dose-response VLPs of wild-type M in the presence of dose-increasing L302A and L305A. Constant amounts of plasmid encoding HA-M were transfected singly or jointly with dose-increasing plasmids encoding Myc-ML302A or Myc-ML305A into 293T cells. At 48 h posttransfection, the cells and medium were harvested, and a VLP budding assay was performed as described in the legend to Fig. 1. WB was performed with anti-HA and anti-Myc Ab, and then the cell lysate blot was stripped and reprobed with an anti-GAPDH Ab as a loading control. Quantification of the budding index for each group is shown. Standard errors were calculated from three independent experiments.