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Site-directed mutagenesis of the carboxy-terminal RGG2 motif affects subcellular localization of the ORF57 protein. 293T cells were transfected with either pGFP, pORF57GFP, or the ORF57 mutants. After 24 h, the cells were fixed and permeabilized, and the GFP fluorescence was analyzed by direct visualization, whereas indirect immunofluorescence was performed using C23-specific antibodies (A) and fibrillarin-specific antibodies (B) to identify the nucleolus.
