Fig. 6
CPEB2 is present in most Slc17a6 mRNA puncta and promotes VGLUT2 synthesis in NMDA/glutamate-stimulated synaptosomes. A DIV17 neurons were probed with Slc17a6 mRNA by FISH, followed by immunostaining of CPEB2 and the axonal marker tau and Hoechst labeling. Representative images showed the colocalization of Slc17a6 mRNA and CPEB2 in somas (defined by bright-field images) and axons (tau+ areas) with magnified images outlined in blue and yellow squares, respectively. Scales, 10 μm and 1 μm in magnified images. B The percentage of Slc17a6+ puncta containing CPEB2 signal in neurons. Twelve image fields from 4 independent cultures were quantified. C Representative images and quantified results showed the number of Slc17a6+ puncta in the SLM layer of CPEB2-cWT and cKONes hippocampi. Scale, 10 μm. Numbers in parentheses (n/N) represent the number of slices (n) and mice (N). D Synaptosomes from adult cortical and hippocampal tissues (8 independent preparations) were stimulated without (Ctrl) or with 50 μM NMDA and 10 μM glutamate for 30 s, followed by the addition of 120 μM APV ± 200 μg/ml cycloheximide (CHX) for the indicated times and then harvested for immunoblotting. Data are mean ± SEM. *P < 0.05 and ***P < 0.001, Mann–Whitney Rank Sum Test in (B), Student’s t test in (C), and two-way ANOVA with Fisher’s LSD post-hoc test in (D)