FIG. 3.
Effect of the NES21 and NES22 substitutions within the NS2 NES site on viral protein synthesis. Lysates of 35S-labeled human 293T and mouse A9 cells, transfected with pdBMVp, pdBMV-NES21, pdBMV-NES22, or no DNA (mock), were precipitated with antisera directed against the NS2 C terminus (A), the NS1 C terminus (B), or the VP1 and VP2 C termini (C). The autoradiograms show immunoprecipitated proteins after separation by electrophoresis through SDS-containing bipartite 8 and 12% polyacrylamide gels. The positions of Crm1 (110 kDa), 14-3-3 (30 and 32 kDa), NS2 (ca. 25 kDa), NS1 (83 kDa), VP1 (83 kDa), and VP2 (65 kDa) proteins are indicated. Molecular masses of prestained standard proteins are shown on the right.