FIG. 5.

Confocal IFA of nsp10 mutants. DBT cells were infected with wt-icMHV and the panel of mutants at an MOI of 5 PFU/cell for 6.5 h and fixed in methanol. Cells were then dual labeled with anti-nsp10 (α-nsp10) and anti-N or anti-M to determine if nsp10 colocalizes with N at sites of replication or with M at sites of viral assembly. (A) In wt-icMHV, nsp10 colocalizes with N predominantly to perinuclear foci, where the viral replication complex is thought to assemble. nsp10 clearly does not colocalize with M, a marker for viral assembly. (B) For the panel of mutants, it appeared that nsp10 localized predominantly at the same perinuclear structures as wt-icMHV, with incidental staining occurring in the nucleus in mutant nsp10-E2.