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. 2023 Jan 21;14:349. doi: 10.1038/s41467-023-35965-8

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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. 8 — a Time course analysis of TJ formation in CTRLsh and GPC4sh hiPSCs after cell dissociation and seeding in combination with the analysis of pSMAD1,5 expression in response to BMP4 stimulation for 2, 6, 9, 12 and 18 h. Cells were subjected to immunofluorescence analysis of pSMAD1,5 (magenta) and ZO1 (green) proteins, n = 3. Scale bar: 100 μm. b Quantitative analysis from the staining shown in a of pSMAD1,5 positive cells (left panel) and of ZO1 organized areas (middle panel) in CTRLsh and in shGPC4 hiPSC cultures. Note the different kinetics in GPC4sh cultures versus controls. The right panel show the combination of the left and middle panels to highlight the correlation between the percentage of pSMAD1,5 positive cells and of ZO1 organized area. Data are represented as mean ± SEM, n = 3 for pSMAD1,5 staining and n = 4 for ZO1 staining. Statistical analysis: Statistical analysis: (b) two-way ANOVA followed by Sidak’s multiple comparison test were used to analyzed pSMAD1,5 counts and unpaired t-test were used to analyzed ZO1 organized areas P-values: (***) <0.001, (*) <0.05. For all panels “n” corresponds to the number of biological replicates unless stated otherwise. Source data are provided as a Source Data file.