ER stress induces EBV lytic genes in LCLs. (A) Schematic representation of drug treatments in LCLs. Cells were treated with DMSO (panel 1) or 40μM SAL (panel 3) for the total length of the experiment; with a 6-hour pulse of 50μM TG and then cultured in media lacking TG for the remaining length of the experiment (panel 2); or with a 6-hour pulse of 50μM TG as in panel 2 but in the presence of 40μM SAL for the total length of the experiment (panel 4). (B) Induction of EBV lytic genes 24 hours after chemical treatment. At 24 hours after drug treatment, LCLs were harvested, RNA isolated, and gene expression of immediate-early (BRLF1 and BZLF1), early (BMRF1), and late (gp350) EBV lytic genes was evaluated by real-time qPCR. Comparison of DMSO versus TG, DMSO versus TG + SAL, and TG versus TG + SAL were all statistically significant at P < .05. (C) Induction of EBV lytic genes 72 hours after drug treatment, when LCLs were harvested and treated as in panel B. Comparison of DMSO versus TG, DMSO versus TG + SAL, and TG versus TG + SAL were all statistically significant at P < .05. (D) TM treatment also induces EBV lytic genes. LCLs were treated with a 4-hour pulse of TM (5 μg/mL) or TM + SAL and then cultured for a total of 72 hours. At 72 hours after drug treatment, LCLs were harvested as described and EBV lytic gene expression was analyzed by real-time qPCR. All comparisons were statistically significant.