Fig 1.

ICP27 mutant WRL-A does not interact with Aly/REF. (A) HeLa cells were cotransfected with pEGFP-Aly/REF and wild-type (WT) ICP27 or plasmids expressing ICP27 mutant WRL-A, D6-21-A, or D6-21-A+WRL-A as indicated. Transfected cells were infected with ICP27 null mutant virus 27Lac-Z 24 h after transfection. Four hours after infection, cells were fixed, and immunofluorescence staining was performed with anti-ICP27 antibody. Green fluorescent protein (GFP) fluorescence was visualized directly. The white arrows show the colocalization of Aly/REF and ICP27. The yellow arrows point to Aly/REF sites that do not colocalize with ICP27 harboring the WRL mutations. (B) HeLa cells were transfected with WT ICP27 or mutant WRL-A or with pUC18 as a negative control. Twenty-four hours after transfection, cells were infected with 27-LacZ. Eight hours after infection, cells were harvested, and immunoprecipitation (IP) was performed on cell lysates with anti-ICP27 antibody (α-ICP27). Western blots (WB) were probed with anti-Aly/REF and anti-ICP27 antibodies.