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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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Characterization of neuronal subtype differentiation and synaptic marker in iPSCs-derived neuronal cells from GD patients. (a) Representative images of TUJ1, S100b, Hoechst and merge fluorescence in iPSCs-derived neuronal cells from control (top), GD3-1 (middle) and GD3-2 (bottom) samples. (b) Summary of the relative expression of TUJ1 over GFAP immunofluorescence per cell in GD3-1 and GD3-2 compared to control group (Kruskal–Wallis test: H(2) = 1.69, p = 0.51, n = 3 technical replicates). (c) Bar graphs representing the relative mRNA expression levels of NeuN (top left, Kruskal–Wallis test: H(2) = 6.02, p = 0.08 for Control vs. GD3-1, p > 0.99 for Control vs. GD3-2, n = 5 technical replicates), VGLUT1 (Kruskal–Wallis test: H(2) = 11.24, p = 0.09 for Control vs. GD3-1, p = 0.09 for Control vs. GD3-2, n = 6 technical replicates), GAD67 (bottom left, Kruskal–Wallis test: H(2) = 8.57, p = 0.01 for Control vs. GD3-1, p = 0.04 for Control vs. GD3-2, n = 6 technical replicates) and GFAP (bottom right, Kruskal–Wallis test: H(2) = 2.21, p = 0.35, n = 6 technical replicates) in iPSCs-derived neuronal cells from control, GD3-1 and GD3-2 samples. (d) Representative images of TUJ1, Synaptophysin, Hoechst and merge fluorescence in iPSCs-derived neuronal cells from control (top), GD3-1 (middle) and GD3-2 (bottom). e. Summary of the relative average synaptophysin immunofluorescence per cell in GD3-1 and GD3-2 compared to control group (Kruskal–Wallis test: H(2) = 9.85, p = 0.02 for Control vs. GD3-1, p < 0.01 for Control vs. GD3-2, n = 5 technical replicates).