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. 2023 May 4;9:144. doi: 10.1038/s41420-023-01445-7

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© The Author(s) 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 4 — A Representative images showing the influences of incubation with 1 nM NPCT and d-NPCT for 24 h on the excitatory synaptic marker PSD-95 and inhibitory synaptic marker gephyrin in primary hippocampal neurons. B–D Quantifications of the density of PSD-95 puncta, gephyrin puncta, and the ratio of PSD-95/gephyrin. One-way ANOVA with Turkey’s post hoc analyses in (B, C) and Kruskal–Wallis H test in (D). n = 28 dendrites from 3 independent experiments in each group. **P < 0.01, ***P < 0.001. E Representative images demonstrating the neuritogenesis effects of 1 nM NPCT and d-NPCT incubation for 24 h in primary hippocampal neurons. F Statistical analysis showing the effects of NPCT on the total dendrite length. Kruskal–Wallis H test with Dunn’s post hoc analyses. n = 25 neurons from 3 independent experiments in each group. ***P < 0.001. G Linear sholl analysis showing that NPCT facilitated dendritic arborization in primary hippocampal neurons. Repeated-measure two-way ANOVA with Dunnett’s post hoc analyses. n = 25 neurons from 3 independent experiments in each group. *P < 0.05, **P < 0.01, ***P < 0.001, compared with control group. H–J Semi-log sholl analyses demonstrating that NPCT increased the complexity of dendritic arborization. A fitting line was generated by simple linear regression for each group and the k coefficient, calculated by multiplying negative one with the slope of the fitting line, was compared between groups. n = 25 neurons from 3 independent experiments in each group. ***P < 0.001, compared between groups. K Representative images showing the spinogenesis effects of NPCT in primary hippocampal neurons. L Statistical analysis demonstrating that NPCT increased the spine density. One-way ANOVA with Turkey’s post hoc analyses. n = 40 dendrites, 52 dendrites, and 44 dendrites from 3 independent experiments in the control, NPCT, and d-NPCT group, respectively. ***P < 0.001. M Immunoblots showing the effects of NPCT on the expression of full-length and cleaved caspase-3. n = 4 independent experiments. All data except (D, F) are presented as mean ± SEM. Data in (D, F) are presented as median with IQR.