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. 2017 Jun 13;3:17019. doi: 10.1038/celldisc.2017.19

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Copyright © 2017 The Author(s)

This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

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UHRF1 expression pattern in the three-dimensional tracheosphere assay. (a) Schematic for the three-dimensional organoid culture system. HBE cells were mixed with fibroblasts in growth factor reduced Matrigel. (b) Representative microscopic images of tracheosphere cultures at days 7, 14 and 28. Single basal cells proliferated and self-renewed to form visible spheres within 1 week, and spheres began to differentiate after 2 weeks. (c) Confocal images of individual spheres collected at indicated time points to show the expression levels of UHRF1, KRT5 and KRT8. (d) Confocal images of individual spheres collected at indicated differentiation time points stained with the UHRF1 antibody and the EdU click-it imaging kit. Spheres were incubated with EdU for 5 h before staining. UHRF1 was expressed in all detectable EdU-positive cells. Scale bars: 20 μm.