Subcellular distribution of AMOT-130 and -80 in hippocampal neurons. (A) Neurons transfected with cDNAs encoding YFP–AMOT-80 and YFP–AMOT-130 (green) were immunostained for endogenous MAP2 (red). (B) Coimmunostaining of endogenous MAP2 (blue) together with Bassoon (Bsn, red; left) or gephyrin (Geph, red; right) in neurons expressing YFP–AMOT-130. (C) Quantification of YFP–AMOT-80 punctate as a fraction of total punctate numbers in soma (So), dendrites (Dd), and branch points (Bp) in culture at indicated stages. *, P < 0.05; one-way ANOVA with Tukey’s post hoc test. (D) Percentage of Bassoon and gephyrin clusters that overlap with YFP–AMOT-130. **, P < 0.01; two-way ANOVA with post hoc Sidak’s multiple comparisons test. (E) Schematic of AMOT-130 mutant constructs used in this study. AB, actin-binding domain. (F) Clusters of YFP–AMOT-130 distributed with mCherry–PSD-95 (left), RFP-actin (middle), or HA-MUPP1 (right). (G) Manders’s colocalization coefficient for clusters of YFP–AMOT-130 that overlap with mCherry–PSD-95 or RFP-actin. Error bars represent means ± SD. *, P < 0.05; Student’s t test. (H) Neurons coexpressing YFP–AMOT-130–ΔC and mCherry–PSD-95. (I) Neurons expressing YFP–AMOT-130N together with mCherry–PSD-95 or RFP-actin. Bars, 5 µm. (J) Overlap of YFP–AMOT-130N and RFP-actin clusters in spines. Pearson’s correlation coefficient = 0.961; P < 0.01.