Fig. 2.

The interaction between vhs and VP22 is mediated by VP16. HEK 293T cells were transiently transfected with plasmids encoding full-length VP22 or full-length VP16 either alone or in combination. Soluble cell extracts were prepared 48 h after transfection, reacted with glutathione–agarose beads bound to GST (lanes 4, 6, and 8) or GST–VHS (lanes 5, 7, and 9). The proteins bound to the beads were subjected to electrophoresis on a denaturing polyacrylamide gel, transferred to a nitrocellulose membrane, and reacted with antibodies against VP16 (upper blot) or VP22 (lower blot). Immunoreactivity of VP22 (lanes 1 and 3) and VP16 (lanes 2 and 3) from a one-tenth volume of whole-cell lysates also is shown. cDNA, pcDNA 3.1(+).