Figure 2

NMDAR-elicited Wnt5a increase requires translation but not transcription. A. Primary cortical neurons (10 DIV) were pre-treated with vehicle (Control) or 20 μΜ anisomycin for 30 min, and then incubated with 50 μΜ NMDA for 15 min, followed by Wnt5a immunoblotting. The Graph is a summary of three independent experiments (**, p < 0.01; One-way ANOVA). B. Primary cortical neurons (10 DIV) were pre-treated with vehicle (Control) or 20 μΜ actinomycin D for 30 min, followed by addition of 50 μΜ NMDA for 15 min. Wnt5a protein was detected by Western blotting and quantified. The Graph is a summary of four independent experiments (*, p < 0.05; **, p < 0.01; One-way ANOVA). C. Primary cortical neurons (10 DIV) were treated with vehicle (Control) or 50 μΜ NMDA for 15 min. Wnt5a mRNA was quantified by Real-time RT-PCR (qPCR). The summary graph is from three independent experiments (40 cycles, CT values: 25.1 ± 0.5/control vs. 25.6 ± 0.3/NMDA; p > 0.05; two-tailed Student's tests). D. Melt curve of Wnt5a qPCR on control cells. The melt curve on NMDA-stimulated cells was similar (not shown). E. RT-PCR results of Wnt5a in control and NMDA-treated cells.