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. 2011 Jan 19;85(7):3356–3366. doi: 10.1128/JVI.02105-10

FIG. 2.

FIG. 2.

ICP0 induces polyubiquitylation of E2FBP1. (A) The level of polyubiquitylated E2FBP1 increased during the immediate-early phase of HSV-1 infection. HEK293FT cells expressing HA-E2FBP1 and His-Ub were infected with HSV-1 at an MOI of 10. Cell lysates were prepared at the indicated times postinfection. The left and middle panels show expression levels of ICP0 (lanes 1 to 3) and HA-E2FBP1 (lanes 4 to 6). The right panel shows polyubiquitylated forms of HA-E2FBP1 (lanes 7 to 9) collected from cell lysates. (B) Polyubiquitylation of E2FBP1 was enhanced by expression of ICP0. The relative ratios of plasmids transformed into HEK293FT cells are shown at the top of the figure. Cell lysates were prepared after 20 h of transformation and subjected to immunoblotting (lanes 1 to 6) or Ni-NTA pulldown (lanes 7 to 12). The positions of ICP0 and nonubiquitylated endogenous or His-tagged E2FBP1 are indicated on the left. mpi, minutes postinfection.