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. 2022 Jun 6;14:890231. doi: 10.3389/fnsyn.2022.890231

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Copyright © 2022 Lutz, Bauer, Ioannidis, Schön and Boeckers.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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ICC analysis in primary rat hippocampal neurons. (A) Neurons have been fixed at DIV14 and secondary dendrites were analyzed. Cells were stained against MAP2, SYNAPSIN1/2, or BASSOON and (B) Rock-GS, (C) SY-311, (D) SY-302, (E) SY-304, (F) ASC, (G) CS, (H) Fr1+2, (I) Fr1+2+3, and (J) vNterm. Scale bar 5 μm. ICC and the derived mask obtained in ImageJ with FindFoci are shown. Left graph: Number of synapses. SHANK3 puncta co-localizing with the respective pre-synaptic marker (red) or not co-localizing (blue) are shown. Right graph: Size of SHANK3 puncta and estimated density. The boxplot shows the median and the interquartile range, the whiskers cover minimal to maximal values. The white dot marks the mean. Data collected from a total of 40 cells of 4 independent experiments. Groups were tested for normality using the Shapiro–Wilk test and then compared using the Wilcoxon test. **p ≤ 0.01, ****p ≤ 0.0001.