Fig. 5. (A) Homological amino acid sequences conserved in HSV UL14 and Hsp70 family. The red letters are complete matches, and the blue letters are amino acids with similar properties. (B) The comparison of the ability of UL14 wild type and mutant proteins to translocate VP26-FLAG in coexpressing cells. Cells were cotransfected with pFLAG-UL35 and the plasmids indicated (pcDNA3-) and, 24 hours later, fixed and stained for the two proteins. Predominant nuclear localization of VP26-FLAG was counted in at least 150 double-stained cells. The bars show the percentage of coexpressing cells with nuclear localization of VP26-FLAG. (C-E) Double-staining of cells transfected with pcDNA3-UL14R (60,64) A and pFLAG-UL35. It is shown that the UL14 mutant protein (C) colocalizes with VP26-FLAG (D) in the cytoplasm but no nuclear staining of VP26-FLAG is observed (E, merge). Images in C-E are representative of VP26 cytoplasmic localization in the assay shown in Fig. 5B.