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. 2024 Apr 22;14:9177. doi: 10.1038/s41598-024-59834-6

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© The Author(s) 2024

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Increase in Caffeine-induced Ca²⁺ release in iPSCs derived neuronal cells from GD. (a) (Left) Maximum intensity projection (MIP) images of Fluor-8 fluorescence of iPSCs-derived neuronal cells from control, GD3-1 and GD3-2 samples. (Right) Two representative traces recorded from cells marked with a respective color in the MIP images corresponding to the left panel during caffeine application (indicated by tick mark). (b) (Left) Maximum intensity projection (MIP) image of Fluor-8 fluorescence of iPSCs-derived neuronal cells from control, GD3-1 and GD3-2 samples pretreated with cyclopiazonic acid for 30 min. (Right) Two representative traces recorded from cells marked with a respective color in the MIP images corresponding left panel during caffeine application (indicated by tick mark). (c) (Left) Histograms representing the distribution of area under the curve of Ca²⁺ transients during caffeine application (blue area in A) in iPSCs-derived neuronal cells from control (black), GD3-1 (orange) and GD3-2 (red) samples. (Right) Cumulative distribution of area under the curve of Ca²⁺ transients during caffeine application (blue area in A) in iPSCs-derived neuronal cells from control (black), GD3-1 (orange) and GD3-2 (red) samples (Two-sample Kolmogorov–Smirnov test: D = 0.33, p < 0.01 for Control vs. GD3-1; D = 0.19, p = 0.02 for Control vs. GD3-1, n = 177, 150 and 83 cells for control, GD3-1 and GD3-2, respectively). (d) Summary of the average area under the curve of Ca²⁺ transients during caffeine application per experimental in control, GD3-1 and GD3-2 groups (Kruskal–Wallis test: H(2) = 8.83, p = 0.02 for Control vs. GD3-1, p > 0.99 for Control vs. GD3-2, n = 9,8 and 8 technical replicates for Control, GD3-1 and GD3-2, respectively). e. Summary of the average area under the curve of Ca²⁺ transients during caffeine application with pretreatment with cyclopiazonic acid per experimental in control, GD3-1 and GD3-2 groups (Kruskal–Wallis test: H(2) = 0.89, p = 0.66, n = 7,7 and 6 technical replicates for Control, GD3-1 and GD3-2, respectively).