Figure 7.
Identification of the interacting domains of VP8 and gM. EBTr cells were co-transfected with FLAG-tagged truncated VP8 versions and pgM-HA or with HA-tagged truncated gM versions and pFLAG-VP8, and the lysates were either directly analyzed by SDS-PAGE (A,B) or incubated with anti-FLAG (C,D) or anti-HA (E) beads. After elution by SDS loading dye, the samples were subjected to SDS-polyacrylamide gel electrophoresis on a 4–15% gel and transferred to 0.45 µm nylon membranes. (A) Expression of truncated versions of VP8 protein. (B) Expression of truncated versions of gM protein. (C) Immunoprecipitation of full-length gM-HA by N-terminal VP8 deletions shows an interaction of gM-HA with all VP8 versions except the deletion lacking amino acids 1–631. (D) Immunoprecipitation of full-length gM-HA by C-terminal VP8 deletions shows an interaction of gM-HA with VP8 consisting of amino acids 1-631, but not with the other VP8 versions. (E) Immunoprecipitation of full- length VP8-FLAG by truncated gM-HA proteins shows an interaction of VP8-FLAG in the presence of the amino acids 70–438, 140–438, and 210–438 of gM-HA. Molecular weight markers × 10−3 are shown in the left margins.