Fig. 8.
Permeabilization activity of 2B-derived constructs in BHKT7 cells. (A) Structural organization of the transfected 2B variants. (B) BHKT7 cells were transfected with pTM1-2Bwt or with pTM1-2B variants. At 3 h posttransfection, cells were pretreated with hygromycin B (HB) at 0.5 mM for 15 min and then labeled with [35S]Met/Cys for 45 min in the presence of the inhibitor. After labeling, the proteins were analyzed by SDS-PAGE (17.5%) followed by fluorography and autoradiography (top). The synthesis of 2B protein was detected by Western blotting using specific rabbit polyclonal antibodies (bottom). The numbers below each lane represent the percentage of protein synthesis in the presence of hygromycin B as calculated by densitometric scanning.