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. 2020 Feb 28;10:3684. doi: 10.1038/s41598-020-60680-5

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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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MET process with epigenetic changes associated with somatic cell reprogramming induced by P334. (A) RT-PCR analysis of MET genes following reprogramming in control and P334 treated after dox induction (data represent mean ± SEM. **p < 0.01). (B) RT-PCR analysis of pluripotent markers Oct3/4, Sox2, Esrrb, Ssea1 and E-cadherin (Cdh1) in OSKM-infected fibroblasts with and without P334 treated at 15 days after dox induction (data represent mean ± SEM. *p < 0.05, **p < 0.01). (C) Western blot analysis of histone 3 lysine 4 tri-methylation (H3K4me3) and NANOG of 4 factors infected fibroblasts reprogramming in control and P334 treated batch. (D) Immunostaining analysis showing the expression of pluripotency markers, SSEA1 and CDH1, in OSKM-infected fibroblasts with and without P334 and noggin (nog) treated at 15 days after dox treated. (Scale bar: 150 um) (E) Immunostaining analysis showing the co-localization of pluripotency markers, SSEA1 and CDH1, in OSKM-infected fibroblasts with and without P334 and nog treated at 15 days after dox treated. More amount of co-localized cells were observed in P334 treated batch. However, lowered amount of co-localized cells were measured in nog treated batch. (White triangle showing the co-localized cells in single iPSC colony.) (Scale bar: 30 um) (F) Bar graph representing the number of CDH1 positive iPSC colonies per field (data represent mean ± SEM. *p < 0.05, **p < 0.01). (G) Bar graph representing the number of co-localized CDH1 and SSEA1 positive cells in single iPSC colony (data represent mean ± SEM. **p < 0.01). (H) QRT-PCR analysis of pluripotency genes (Oct3/4, Sox2 and Nanog) in OSKM-infected fibroblasts with and without P334 and nog treated (data represent mean ± SEM. *p < 0.05, **p < 0.01). Data is analyzed at 15 days after dox induction.