FIG. 9.

Isolation of stable HBV core protein-SRPK2 complexes from HuH-7 cells. (A) HuH-7 cells were infected in six-well dishes with adenovirus at 5,000 particles of AdHBVcore per cell plus either control adenovirus (no expression cassette), 25,000 particles of AdSRPK1-VSV per cell, or 15,000 particles of AdSRPK2-VSV per cell as indicated. Control adenovirus was added to give a total particle number of 30,000/cell for each of the infections. On the second day after infection, cell lysates were prepared and subjected to immunoprecipitation (IP) with polyclonal anti-HBV core protein antibody. Samples were then resolved by SDS-13% PAGE and analyzed by immunoblotting with anti-VSV antibody (upper panel) or anti-HBV core protein antibody (middle panel). In parallel, total cell lysates were analyzed by immunoblotting with anti-VSV antibody (lower panel). Positions of SRPKs and HBV core protein are indicated on the right. (B) HuH-7 cells were transiently transfected in 10-cm dishes with control plasmid, replication-competent HBV plasmid pSPT1.2xHBV (15 μg/dish), or plasmid encoding SRPK2-VSV (5 μg/dish), as indicated. After 48 h, cell lysates were prepared and processed as described for panel A. In addition, total cell lysates were analyzed by immunoblotting with anti-HBsAg antibody (lower panel). Positions of SRPK2, HBV core protein, and the different forms of HBsAg are indicated on the right.