FIGURE 5.

R(+)WIN55,212-2 targets JNK to promote IFN-β expression. A and B, primary mouse astrocytes were seeded into 6-well plates and treated with R(+)WIN55,212-2 (20 μm) (A) or fenofibrate (20 μm) (B) for various times (5–240 min). Cell lysates were subsequently subjected to Western immunoblotting using anti-phospho-JNK, anti-total JNK, and anti-β-actin antibodies. C, primary astrocytes were treated with GW6471 (1 μm; 1 h) prior to R(+)WIN55,212-2 (20 μm) for 15 min, and lysates were prepared and subjected to immunoblotting. All immunoblots were subjected to densitometric analysis with levels of phospho-JNK normalized to total levels of JNK. D and E, HEK293-TLR3 cells were co-transfected with IFN-β promoter-regulated firefly luciferase and TK Renilla luciferase, left overnight, and pretreated (1 h) with SP600125 (10 μm) in the absence or presence of R(+)WIN55,212-2 (20 μm) (D) or fenofibrate (20 μm) (E) prior to poly(I:C) (25 μg/ml) for 6 h. Lysates were assayed for firefly luciferase activity. Data are mean ± S.E. (error bars) of triplicate determinations and are representative of three independent experiments (D and E) or represent densitometic data from 5–9 animals (A–C). *, p < 0.05; **, p < 0.01; ***, p < 0.001 compared with vehicle-treated cells. +, p < 0.05; ++, p < 0.01; +++, p < 0.001 compared with R(+)WIN55,212-2-treated cells (C) or poly(I:C)-treated cells (D and E). $, p < 0.05 compared with cells treated with poly(I:C) in the presence of R(+)WIN55,212-2 or fenofibrate.