Fig. 5
Nlg1 C-terminal truncation affects AMPAR-dependent synaptic transmission and spine density. a Diagram of Nlg1 truncation mutants lacking the last 5 aa comprising the C-terminal PDZ domain-binding motif (Nlg1-Δ5) or the last 72 amino acids also including the gephyrin-binding motif (Nlg1-Δ72). TM: transmembrane domain. b Representative traces of evoked AMPAR- and NMDAR-mediated EPSCs recorded at −70 mV and +40 mV, respectively. Color sample traces correspond to neurons co-electroporated with GFP and Nlg1-WT, Nlg1-Δ5, or Nlg1-Δ72, respectively, while black traces correspond to control, unelectroporated neurons. c, d Average AMPAR- and NMDAR-mediated EPSC amplitudes, respectively, normalized to control (the dashed line indicates 100%, dot plots corresponds to different pairs, from 3 independent experiments). e Confocal images showing CA1 neurons from Nlg1 KO organotypic slices electroporated with GFP (green), BirAER, and AP-tagged Nlg1-Δ5 or Nlg1-Δ72 which were labeled with streptavidin-Atto647 (red). Scale bars, 30 µm (upper panels) and 10 µm (lower panels). f Average spine density for CA1 neurons electroporated with EV, Nlg1-WT, Nlg1-Δ5, or Nlg1-Δ72 (dot plots corresponds to different cells, from 2 independent experiments). Data in graphs c and d were compared to the control condition by Wilcoxon matched-pairs signed rank test, and between themselves using one-way ANOVA followed by Tukey’s multiple comparison (ns: not significant, *P < 0.05, ***P < 0.001). Data in graph f were compared by a Kruskal–Wallis test followed by Dunn’s multiple comparison test (**P < 0.01). Data represent mean ± SEM
