FIG. 11.

Coimmunoprecipitation of pU_L17 and pU_L25 in the absence of other viral proteins. Hep2 cells were transfected with the indicated pU_L25 expression plasmids or pcDNA3. Lysates of the transfected cells were prepared 24 h later and reacted with purified pU_L17 fused at the C terminus with a histidine tag. After 2 h, the lysates were prepared and precleared. They were then reacted with pU_L25-specific antibody, and immune complexes were purified, denatured, and electrophoretically separated on a denaturing polyacrylamide gel. The proteins were transferred to nitrocellulose and probed with pU_L25- and pU_L17-specific antibodies. (A) Immunoblot probed with pU_L25-specific antibody. Lanes 1 to 4, total cellular lysates of Hep2 cells transfected with the indicated plasmids; lanes 5 to 9, proteins reacted with purified pU_L17-His and immunoprecipitated with pU_L25-specific antibody. The nature of the relevant protein expressed in each sample is indicated above each lane. (B) Immunoblot probed with pU_L17-specific antibody. The nitrocellulose sheet used in panel A was stripped of bound immunoglobulin and probed with pU_L17-specific antibody.