Fig. 6. Western analysis of E1B 55-kDa expression in KB cells. KB cells were mock-transduced or transduced with the viruses indicated below each lane (genotypes are presented below) and harvested 24 h later, and the cells were lysed in SDS protein sample buffer [60 mM Tris·Cl, pH 6.8/1% SDS/5 mM EDTA/50 mM DTT/10% (vol/vol) glycerol) at 2 x 10⁷ cells per ml. The cells were disrupted by sonication, and the lysates were clarified by centrifugation before separating the proteins from 5 x 10⁴ infected cells per lane by SDS/PAGE through a 12.5% acrylamide gel. The separated proteins were electrophoretically transferred to a nitrocellulose support and visualized by blotting with the E1B-55-kDa-specific mouse mAb, 2A6 (1) followed by chemiluminescent detection using the PicoWest kit as per the manufacturer’s recommendations (Pierce.). The positions of molecular weight markers are indicated to the right.

1. Sarnow, P. Sullivan, C. A. & Levine, A. J. (1982) Virology 120, 510-517.