FIGURE 10.

IQSEC3 is required for maintenance of gephyrin clusters in cultured neurons. A, representative images of both dendrites and soma of cultured hippocampal neurons transfected at DIV8 with lentiviral constructs expressing EGFP alone (Control), co-expressing EGFP with shRNAs against Iqsec3 (IQSEC3-KD), or co-transfected with IQSEC3-KD and the shRNA-resistant IQSEC3 full-length vector (+IQSEC3 rescue). Neurons were analyzed by double immunofluorescence labeling for gephyrin (red) and EGFP (blue) at DIV14. Scale bar, 10 μm (applies to all images). B, summary graphs of the effects of IQSEC3 KD in neurons on gephyrin puncta density (left), gephyrin puncta size (middle), and gephyrin puncta intensity (right). At least five to eight dendrites or one soma per transfected neuron were analyzed and group-averaged. Data are presented as means ± S.E. (*, p < 0.05; ³*, p < 0.001; ANOVA with Tukey's test). More than 850 gephyrin puncta were quantified in an individual image. AU, arbitrary units.