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. 2016 May 19;7(5):e2227. doi: 10.1038/cddis.2016.118

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Copyright © 2016 Macmillan Publishers Limited

Cell Death and Disease is an open-access journal published by Nature Publishing Group. 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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Rescue of corneal epithelial differentiation of EEC-iPSCs by mutated-p63 allele-specific silencing RNAs. WT-iPSCs and EEC-iPSCs were subjected to corneal epithelial differentiation protocol. Phase contrast pictures of WT-iPSCs and EEC-iPSCs at days 0, 10 and 13 of differentiation are shown in a. Cells were harvested at days 13–15 of differentiation and subjected to RT-qPCR analysis of the indicated genes (b), flow cytometry analysis (c) and immunofluorescence staining (d) of K14. EEC-iPSCs were transfected with nonspecific control silencing RNA (SCR) or specific siRNAs against p63 mutated allele (T4 and T11) at days 8–10 during differentiation into corneal epithelial cells. Cells were harvested at day 13–15 and subjected to RT-qPCR analysis of the indicated genes (e) and flow cytometry analysis of K14 (f). Gene expression was normalized to GAPDH housekeeping gene (b and e) and data represent the fold change in gene expression compared with undifferentiated cells (b) or compared with SCR (e). Flow cytometry data represent the percentage of K14+ cells in the cell population (c) and the relative increase in the K14+ cells compared with SCR (f). Results are presented as mean±S.D. (Scale bar, 100 μm)