FIG 6.

Interferon and Wnt pathways intersect through direct GSK3β/TBK1 interaction. (A) miR-Ct/-34a-transfected HeLa cells were mock or SeV treated for 6 h. Total protein was assessed for phosphorylated TBK1 and β-actin levels by Western blotting. Results are representative of data from at least three independent experiments. (B) miR-Ct/-34a-transfected cells were transfected with a plasmid encoding Flag-tagged TBK1 at 24 h post-miRNA transfection. At 24 h post-plasmid transfection, cells were treated with SeV for 6 h, lysed, and subjected to immunoprecipitation with Flag-Sepharose beads. Immunoprecipitated and total fractions were analyzed by Western blotting for GSK3β levels. IB, immunoblotting. (C) miR-Ct/-34a-transfected cells were treated with SeV as described above, with or without the GSK3β inhibitor SB216763 (5 μM). Phosphorylated and total IRF3 levels were assessed by Western blotting. Results are representative of data from at least three independent experiments. Fold changes were calculated by measuring the integrated density using ImageJ software and normalized to the values for the control.