FIG. 5.

A26-A27 protein complexes formed prior to virion assembly and did not require A17 protein. (A) Immunoblot analyses (IB) of 70-/90-kDa complex in the infected cells using anti-A27 Ab. BSC40 cells were infected with individual vaccinia viruses as shown at the top at an MOI of 10 PFU per cell. Wt, wild-type vaccinia virus (WR). Cell lysates were boiled and separated by SDS-PAGE on nonreducing (without 2ME) gels and probed with anti-A27 Abs. The bottom portion shows a longer exposure of a portion of the blot (marked by the dotted line). The black and white arrows indicate the 90- and 70-kDa protein complexes, respectively. A27-m, A27-d, and A27-t represent monomers, dimers, and trimers of A27 protein, respectively. (B) Samples were treated with reducing condition (with 2ME) and loaded in the same order as that used for panel A and were probed with anti-A27 Ab. (C) Samples were loaded in the same order as that for panel A and were treated with reducing conditions (with 2ME) and probed with anti-A17 Ab. (D) Samples were loaded in the same order as that used for panel A and were probed with anti-A26 Ab. A26 red, reduced form of A26 protein; A26 ox, oxidized form of A26 protein; and A26t, truncated A26 protein (aa 1 to 426) (5); M, mock-infected cells; Wt, wild type virus.