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. 2007 Mar 14;81(10):5166–5180. doi: 10.1128/JVI.00120-07

FIG. 8.

FIG. 8.

MHV68 ORF36 kinase induces chromosome condensation in transiently transfected HeLa cells. (A) Slide-cultured HeLa cells were transfected with Flag-tagged HSV-1 UL13-, HCMV UL97-, or MHV68 ORF36-expressing plasmids or control vector. At 24 h posttransfection, cells were fixed and stained for Flag-tagged protein as described in Materials and Methods. Cellular DNA was stained with Hoechst 33258. Flag-tagged protein-positive cells (∼200 cells/set) with condensed chromosomes were counted. Chromosome condensation was observed in 93.2% of cells expressing Flag-tagged MHV68 ORF36. (B) Flag-tagged protein expression was confirmed by immunoblotting using anti-Flag antibodies. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) served as a loading control. Numbers at left are molecular masses in kilodaltons. (C) Phylogenetic analysis of HSV-1 UL13, HCMV UL97, EBV BGLF4, Kaposi's sarcoma-associated herpesvirus (KSHV) ORF36, and MHV68 ORF36. PAM (point-accepted mutations), 1 unit of evolution as the amount of evolution that will change 1 in 100 amino acids on average.