FIG 2.

Cell lines expressing Vps4A-EQ inhibit production of HSV and PRV infectious particles. (A, B) Cells expressing tetracycline-inducible Vps4A or Vps4A-EQ were incubated with tetracycline from either 16 h prior to infection (Pre Tet+) or at the time of infection (Pre Tet-). They were then infected with HSV (A) or PRV (B) at a multiplicity of 10 for 1 h at 37°C, acid washed to inactivate unpenetrated virus, incubated in the continued presence of tetracycline for the times shown, and harvested for determination of the titers. Plotted values are means and standard deviations for duplicate (A) and triplicate (B) dishes each titrated in duplicate. (C, D) Cells were grown and infected by HSV (C) or PRV (D) similarly to the experiments whose results are shown in panels A and B except that PNS was prepared at 16 h postinfection, subjected to SDS-PAGE, and Western blotted for HSV-VP5, mRFP1, or a 75-kDa PRV structural protein as indicated. For panels C and D, each pair of lanes corresponds to Vps4A and Vps4A-EQ cells, respectively, under the following conditions: lanes 1 and 2, infected, no Tet preinduction (Pre Tet-); lanes 3 and 4, uninfected, with Tet preinduction (Pre Tet+); lanes 5 and 6, infected, with Tet preinduction (Pre Tet+).