Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2021 Sep 29;41(39):8134–8149. doi: 10.1523/JNEUROSCI.1930-20.2021

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

Copyright © 2021 the authors

SfN exclusive license.

PMC Copyright notice

Figure 3. — Cultured TSC2^−/− neurons exhibited synchronous neuronal activity. A, Ca²⁺ dynamics of iPS-derived TSC2^+/+, TSC2^+/−, and TSC2^−/− neurons at 33 d. Fluorescence images of Fluo-8 and the ΔF/F₀ changes in Fluo-8 signals are shown. Images were taken at 1 Hz. Thirty-six frames of ΔF/F₀ changes in Fluo-8 are presented. B, Raster plots of Ca²⁺ spikes in TSC2^+/+, TSC2^+/−, and TSC2^−/− neurons. Vertical axis represents the individual cells analyzed (∼1 in 70 cells). C, Developmental change in the percentage of experiments in which synchronous Ca²⁺ spikes were observed in TSC2^−/− neurons. Each value was obtained from 6 to 29 independent experiments. D, Percentages of neurons exhibiting spontaneous Ca²⁺ spikes in TSC2^+/+, TSC2^+/−, and TSC2^−/− neurons on various days of culture. Data are mean ± SD. The experimental numbers were 9-29 for each group. Day 8: one-way ANOVA, F_(2,23) = 1.258, p = 0.296. Day 20: one-way ANOVA, F_(2,24) = 2.042, p = 0.152. Day 30: one-way ANOVA, F_(2,49) = 2.029, p = 0.142. Day 40: one-way ANOVA, F_(2,57) = 2.031, p = 0.141. Day 60: one-way ANOVA, F_(2,75) = 2.034, p = 0.138. Day 80: one-way ANOVA, F_(2,34) = 7.907, p = 0.00151. Bonferroni's multiple comparisons test, **p = 0.0013, TSC2^+/+ (n = 13) versus TSC2^−/− (n = 12); *p = 0.0324, TSC2^+/− (n = 12) versus TSC2^−/− (n = 12). Day 100: one-way ANOVA, F_(2,26) = 1.98, p = 0.158. E, Frequencies of Ca²⁺ spikes in TSC2^+/+, TSC2^+/−, and TSC2^−/− neurons on various culture days of neuronal differentiation. The experimental numbers were 9-29 for each group. Day 8: one-way ANOVA, F_(2,23) = 1.517, p= 0.24. Day 20: Kruskal–Wallis test, p = 0.001084, Steel–Dwass test, **p= 0.004896, TSC2^+/+ (n = 9) versus TSC2^−/− (n = 9); **p = 0.004896, TSC2^+/− (n = 9) versus TSC2^−/− (n = 9). Day 30: Kruskal–Wallis test, p = 0.0968. Day 40: one-way ANOVA, F_(2,57) = 7.323, p = 0.00148, Bonferroni's multiple comparisons test, **p =0.0036, TSC2^+/+ (n = 19) versus TSC2^−/− (n = 19); **p = 0.0086, TSC2^+/− (n = 19) versus TSC2^−/− (n = 19). Day 60: Kruskal–Wallis test, p = 0.6337. Day 80: Kruskal–Wallis test, p = 0.0299, Steel test compared with TSC2^−/−, *p = 0.03611, TSC2^+/+ (n = 13) versus TSC2^−/− (n = 12). Day 100: one-way ANOVA, F_(2,23) = 2.644, p = 0.0926. F, Difference in Ca²⁺ spike frequencies between synchronized (synchro) and nonsynchronized (non) TSC2^−/− neurons at 30 and 60 d. Data are mean ± SD. Day 30: Mann–Whitney U test, ***p = 0.00057. Day 60: Mann–Whitney U test, ****p = 0.0000105. The experimental number was synchronized (n = 9) and nonsynchronized (n = 9) at 30 d. The experimental number was synchronized (n = 12) and nonsynchronized (n = 17) at 60 d.