Fig. 4.

Regulation of cell-signaling components on activation of TLR3. (A) Cytokines: C57BL/6 mice were injected i.p. with saline or polyI:C (5 mg/kg), and livers were harvested at 1 hour (n = 5 or 6/group). RNA was isolated from the livers, and mRNA levels of IL-1β, IL-6, and TNFα were determined by real-time PCR analysis as described earlier. (B) Nuclear receptors: C57BL/6 mice were injected i.p. with saline or polyI:C (5 mg/kg), and livers were harvested at 8 and 16 hours (n = 5 or 6/group). RNA was isolated from the livers, and mRNA levels of CAR, PXR, and RXRα were determined by real-time PCR analysis as described earlier. All data are presented as ±S.D. and standardized for cyclophilin mRNA levels. Expression in saline-treated mice was set to 1, and the -fold change after polyI:C treatment was compared with that of the saline-treated controls. *Significant difference (P < 0.05) between saline and polyI:C groups. The experiments were repeated three times. (C) Cell-signaling pathways: Whole-cell extracts were prepared from the livers of saline and polyI:C-treated mice at 1 hour, and the samples were analyzed by immunoblotting. The phosphorylated forms of JNK (P-JNK) and degradation of IkBα were measured as markers of JNK and NF-κB activation, respectively. The experiments were repeated three times.