Figure 2.

The Effect of Increased CYFIP1 and CYFIP2 Gene Dosage on Inhibitory Synaptic Structure

(A) Representative confocal images of hippocampal neurons transfected with CYFIP1^GFP, CYFIP2^GFP, or GFP control for 4 days before labeling at DIV14 with gephyrin antibodies. Scale bar, 2 μm.

(B) Gephyrin cluster analysis showing a significant decrease in gephyrin cluster number and area upon CYFIP1^GFP or CYFIP2^GFP overexpression (cluster number: from 9.9 ± 0.6 to 6.3 ± 0.9 for CYFIP1 and 6.6 ± 0.6 for CYFIP2; cluster area: from 3.1 ± 0.2 μm² to 2.1 ± 0.3 μm² for CYFIP1 and 1.9 ± 0.2 μm² for CYFIP2; n = 20 cells from 4 preparations; Kruskal-Wallis one-way ANOVA, Dunn’s post hoc multiple comparisons).

(C) Representative confocal images of hippocampal neurons transfected as in (A) and surface labeled with GABA_AR-γ2 subunit antibodies. Scale bar, 2 μm.

(D) Cluster analysis of GABA_AR-γ2 surface puncta showing a decrease in cluster number and area upon CYFIP1^GFP or CYFIP2^GFP overexpression (cluster number: from 8.1 ± 0.9 to 5.4 ± 0.8 for CYFIP1 and 5.2 ± 0.7 for CYFIP2; cluster area: from 2.8 ± 0.6 μm² to 1.1 ± 0.2 μm² for CYFIP1 and 1.2 ± 0.2 μm² for CYFIP2; n = 25 cells from 4 preparations; Kruskal-Wallis one-way ANOVA, Dunn’s post hoc multiple comparisons).

^∗p < 0.05; ^∗∗p < 0.01. Bars indicate mean, and error bars indicate SEM.