FIG 8.

Inhibition of BDNF/TrkB signaling decreases downstream phosphorylation of GSK-3β. (A) The Western blot in the upper panel shows detection of serine 9 phosphorylation of GSK-3β in the indicated cell lines treated with DMSO, ANA-12 (20 μM), or K-252a (100 nM). Cells were cultured with 0.5% FBS for 24 h and then received the indicated drug treatments for 6 h. The lower panel shows the same membrane stripped and then probed for actin. (B) ANA-12 treatment restores GSK-3β activity. C10/MJ cells were transfected with p7LEF-Fos-Luc and pRL-SV40-Luc (internal standard), cultured in 0.5% serum, and treated with DMSO or 20 μM ANA-12. The graph shows data from four independent transfection assays, with values showing the average luminescence normalized to that of the DMSO treatment (set to 100). **, P < 0.005 (two-tailed Student t test).