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. 2021 Nov 11;12:6504. doi: 10.1038/s41467-021-26764-0

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© The Author(s) 2021

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. 6 — a Representative time lapse image of calcium influx in GCaMP hES derived cholangiocyte cyst (3D) in response to ATP. Scale bar represents 200 μm. b Representative traces from Fig. 6a showing the intra cellular calcium release in GCaMP hESC-derived cholangiocytes in the response to ATP. c Quantification of maximum fluorescent intensity representing intra cellular Ca²⁺ release in GCaMP hESC-derived hepatoblasts (HB) and cholangiocytes (chol) in the response to ATP, ^*p ≤ 0.05 two-tailed Student’s t test (n = 4 biologically independent samples from HB, n = 3 biologically independent samples from chol). d Quantification of maximum fluorescent intensity representing intra cellular Ca²⁺ release in GCaMP hESC-derived cholangiocytes in the response to different concentrations of TUDCA, ^**p ≤ 0.01, one-way ANOVA (n = 3 biologically independent experiments per each concentration). Data are represented as mean ± SEM. e Representative trace of the intra cellular calcium release in GCaMP hESC-derived cholangiocytes induced by flow in the absence of ATP. f Quantification of maximum fluorescent intensity in GCaMP hESC-CF01derived cholangiocytes with increased flow rate (n = 3). Two-tailed pearson correlation coefficient was calculated by Prism8. g Quantification of maximum fluorescent intensity representing intra cellular Ca²⁺ release in indicated GCaMP derived hepatoblasts (HB), non-ciliated cholangiocyte (cilia-), and ciliated cholangiocytes (cilia + ) in the response to flow, ^****p ≤ 0.0001 one-way ANOVA (n = 3 biologically independent experiments per each group). h Quantification of maximum fluorescent intensity representing intra cellular Ca²⁺ release in indicated hPSC-derived hepatoblasts (HB) and cholangiocytes (chol) in the response to flow, ^**p ≤ 0.01, ^****p ≤ 0.0001 two-tailed Student’s t test (n = 3 biologically independent experiments per each group). Each circle represents the structure measured for intracellular calcium release.