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. 2020 Feb 26;10:3474. doi: 10.1038/s41598-020-60269-y

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© The Author(s) 2020

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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GRP78 is important for reprogramming and is expressed on the surface of iPSCs. (A) Human keratinocytes were retrovirally infected with OCT4, SOX2, KLF4 and c-MYC (OSKM) either alone (4F) or in the presence of shGRP78 or shScramble control. Resulting colonies (~20 days after infection) were stained for Nanog and colony numbers determined relative to the control. (B) Keratinocytes were retrovirally infected with OSKM (4F) or (C) with OSK (3F), in addition to a retrovirus expressing GRP78 or a GFP control. The number of Nanog positive colonies are shown relative to the control. (D) Keratinocytes were retrovirally infected with 4F following a 2-day prime with either a retrovirus expressing GRP78 or a GFP control. Nanog positive colony numbers are shown relative to the control. (E) iPSCs derived from fibroblasts (FiPS4F5) were examined by immunofluorescence for GRP78 (red), and E-cadherin (green, a cell surface marker), and both markers were found to colocalize together (Merge, Inset; arrows). 4,6-Diamidino-2-phenylindole (DAPI) staining shows nuclei. Results were quantified from triplicate samples, and are representative of at least three independent experiments. Error bars depict the standard error mean (SEM). A one-way ANOVA with a Tukey post hoc test (for multiple comparison tests) or an unpaired t-test (when comparing two samples) were used to determine statistical significance compared to controls, with p < 0.05 being considered statistically significant. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001; ns = not significant.