(A) Chemical structures of various ITC used in this study. (B) and (C) HEK293-TLR3 cells (Wt11) were treated with ITC for 1h, then stimulated with 50 μg/mL p(I):p(C) for another 16 h. Cells were harvested for RNA extraction, and total RNA were analyzed by quantitative RT-PCR for ISG56 (B) and IFNβ (C). (D) HEK293-TLR3 cells were treated with the different ITC including PEITC, ERU, ERY and SFN at the indicated concentrations for 1h, then stimulated with 50μg/mL p(I):p(C) for another 6 h. Cell lysates were immunoblotted with anti-ISG56, ISG60, TLR3 and actin antibodies. NS is non-specific band. * p < 0.05 and ** p < 0.01, the ITC treated samples versus p(I):p(C) control sample in Panels B and C performed by one-way ANOVA analysis.