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. 2005 Jan;79(1):597–601. doi: 10.1128/JVI.79.1.597-601.2005

FIG. 2.

FIG. 2.

FIG. 2.

p28-dependent accumulation of ubiquitin in viral replication factories. (A) Colocalization with HA-ubiquitin. HeLa cells were infected with VV-WR, transfected with HA-ubiquitin, and cotransfected with either FLAG-p28, FLAG-p28(C173S/C176S), FLAG-p28(1-204), FLAG-p28(1-184), or EGFP. Cells were fixed and stained as described in the text to visualize HA-ubiquitin, FLAG constructs, EGFP, and I3L. All p28 constructs tested localized to virus factories, although FLAG-p28(1-184) was also cytoplasmic. EGFP was cytoplasmic and appeared to be excluded from the factories. HA-ubiquitin was excluded from viral factories in the absence of p28, while in the presence of FLAG-p28, FLAG-p28(C173S/C176S), or FLAG-p28(1-204), HA-ubiquitin was enriched in virus factories. In contrast, HA-ubiquitin was excluded from viral factories upon transfection with FLAG-p28(1-184). (B) Colocalization with endogenous ubiquitin. HeLa cells were infected with VV-WR FLAG-p28 or VV-WR for 16 h and stained with anti-ubiquitin antibody FK2 and anti-I3L. Ubi, ubiquitin.