Figure 5.

C/EBPβ modulates EBV lytic gene expression and replication by ER stressors. EBV⁺ Akata cells and Rael were treated with 20nM bortezomib (BZ), 1μM thapsigargin (TG), or 2 μg/mL tunicamycin (TU) for 24 hours. (A) RNA was isolated and subjected to real-time qPCR using primers for ZTA. Primers for β-actin were used as an internal control. (B) Viral DNA was isolated and EBV viral load was measured using real-time PCR. Primers to the EBV BamW region were used to quantify EBV copy number. Primers to β-actin were used as an internal control. (C) EBV⁺ Akata and Rael cells were treated with 1μM thapsigargin (TG) and harvested after 24 hours. Western blots were probed for C/EBPβ, CHOP10, and ZTA. (D) C/EBPβ knock-down was performed in EBV⁺ Akata cells using an inducible vector in which the shRNAmir is only expressed in the presence of doxycycline (DOX). To induce shRNAmir expression, 1 μg/mL doxycycline was added to the cells for 72 hours. Cells were treated with 1μM thapsigargin (TG), and RNA was isolated after 24 hours and subjected to real-time qPCR with primers for ZTA. Primers to β-actin were used as an internal control. Error bars indicate SEM.