FIGURE 1.

WDR82 inhibits SeV-induced ISRE activation and IFNB transcription. (A) WDR82 overexpression inhibits activated ISRE report gene. HEK293 cells (1 × 10⁵) were transfected with ISRE reporter plasmid (0.1 μg) together with control (1 μg) or WDR82 plasmid (1 μg). Twelve hours later, cells were infected with SeV and luciferase assays were performed. Lysates were immunoblotted with anti-HA and anti–β-actin. (B) WDR82 inhibits ISRE activation in a dose-dependent manner. HEK293 cells (1 × 10⁵) were cotransfected with ISRE reporter (0.1 μg) and the indicated amounts of WDR82 plasmid, together with control (0.2 μg) or VISA plasmid (0.2 μg). Cell lysates were performed with luciferase assays 24 h after transfection. (C) WDR82 inhibits IFNB gene transcription. WDR82 stably expressed HEK293 cells, and control cells were infected with SeV for 12 h and RT-PCR was performed with IFNB primers. Cell lines were also immunoblotted with anti-Flag and anti–β-actin. (D) WDR82 knockdown promotes IFNB transcription. HEK293 cells were transfected with WDR82 and with three different siRNAs. Forty-eight hours later, cells were infected with SeV for 12 h before performing RT-PCR experiments. The efficiency of WDR82 knockdown was measured with RT-PCR. (E) HEK293 cells were transfected with WDR82 siRNA and treated with TNF-α. The mRNA levels of IκBα and cIAP2 were measured by quantitative PCR. *p < 0.05, **p < 0.01.