FIGURE 3.

Co-stimulation with BDNF and Fsk causes proteasome-dependent EF2K degradation in cortical neurons. A, cortical neurons (14–15 DIV) were treated with Fsk (10 μm) plus BDNF (100 ng/ml) with or without MG132 (25 μm) for 3 or 6 h and EF2K and GAPDH levels in lysates were assessed by SDS-PAGE and immunoblot. B, graph of summary data. A one-way ANOVA with Tukey's post-hoc tests showed that BDNF and Fsk co-stimulation for 3 or 6 h reduced EF2K levels relative to control (3 h: 86.00 ± 3.39, n = 5; 6 h: 72.23 ± 2.548, n = 7, ** or *** p < 0.01 or p < 0.001, respectively, relative to control). Addition of MG132 significantly blocked this effect (3 h+MG132: 99.26 ± 0.67, n = 4; 6 h+MG132: 86.79 ± 3.48, n = 4, # or ##, p < 0.05 or p < 0.01, respectively, relative to stimulation without MG132).