GFI1 mRNA and protein are degraded rapidly during HCMV infection and their degradation is followed by upregulation of EZH2, JARID2 and NDY1/KDM2B and downregulation of JMJD3.

<p><b>A</b>. The GFI1 mRNA levels in lysates of HFFs harvested at the indicated time points after HCMV infection were monitored by real time RT-PCR. <b>B</b>. The GFI1 protein levels in the same cells were monitored by western blotting. <b>C</b>. HCMV infection promotes the degradation of GFI1 at the RNA level. HFFs were mock-infected or infected with HCMV after treatment with Actinomycin D (5 µg/ml). GFI1 mRNA levels in cell lysates harvested at the indicated time points after exposure to the virus were measured by real time RT-PCR. <b>D</b>. Proteasomal inhibition blocks the rapid downregulation of GFI1 in HCMV-infected cells. HFFs were mock-infected or infected with HCMV after pretreatment with MG132 (10 µM). GFI1 protein levels in cell lysates harvested at the indicated time points after exposure to the virus were measured by western blotting. <b>E</b>. GFI1 protein levels in HFFs transduced with the indicated constructs and harvested before, and 2 hours after HCMV infection, were measured by western blotting. <b>F</b>. The expression of EZH2, NDY1/KDM2B, JARID2 and JMJD3 was measured by real time RT-PCR in lysates of HFFs, harvested at the indicated time points after HCMV infection.