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. 2017 Jul 24;7:6294. doi: 10.1038/s41598-017-06546-9

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© The Author(s) 2017

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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Ultra structure and maturation. (A) Electron microscopy image of E-iPSC-HLCs. BC = bile canaliculi; L = lipid; CCP = clathrin coated pits; M = mitochondrion; RER = rough endoplasmic reticulum; N = nucleus; arrow = tight junctions; arrowhead = microvilli. (B) Bright field microscopy of HLCs. Bi-nuleated cells are marked by arrow-heads. (C) Immunofluorescence-based staining of HNF4α in end-stage HLCs. 79.4% (+/−3.7%) of the cells counted positive for HNF4α. Scale bar: 200 μm Alexa Flour 594 (red). (D) Immunofluorescence-based double staining of Albumin (ALB) and HNF4α in end-stage HLCs. 91.7% (+/− 3.9) of HNF4α positive cells were also positive for ALB. Scale bar: 100 μm, Alexa Flour 594 (red) and Alexa Flour 488 (green).