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. 2020 Dec 15;10:21950. doi: 10.1038/s41598-020-78932-9

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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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Generated iPSCs that showed different reprogramming capacity exhibited similar pluripotent characteristics. (A) Represents the ESC-like morphology of the established iPSCs in comparison to H1 and IMR90-1 cells which was observed by light microscopy using 2 different magnifications (× 4 and × 20). (B) Represents the alkaline phosphatase expression that was expressed by the established iPSC colonies compared to that expressed by H1 and IMR90-1 cells. Microscopic images were taken using × 10 magnification. (C) Representative graphs for RT-qPCR analysis of pluripotent genes that were expressed by the generated iPSCs and those expressed levels were compared with reference expressions represented by H1 and IMR90-1. (D) Representative graphs for expression levels of SeV-derived genes. Each graph represents one of the reprogrammed samples (HC, ID-PD, A53T-PD1 and A53T-PD2). Expression levels of pluripotent genes and SeV-derived genes were normalized by the expression of GAPDH as an endogenous control gene and the original fibroblasts were used as negative control for detected genes. Fibro: fibroblasts; Trans. fibro.: transduced fibroblasts on day 7 post transduction. (E) Represents Western blot analysis that shows the expression levels of pluripotent proteins [Oct4 (~ 45 KDa), Sox2 (~ 34 KDa), Nanog (~ 40 KDa), and Klf4 (~ 65 KDa)] that were expressed in the established iPSC colonies and in reference cells H1 and IMR90-1. Left panel shows the proteins expressed in HC and ID-PD colonies; while the right panel indicates the proteins expressed in A53T-PD1 and A53T-PD2 colonies. Gapdh was used as a loading control. Full-length blots are presented in Supplementary Fig. 2. (F) Representative immunofluorescence microscope images of pluripotent markers that were expressed by the generated iPSCs. [Oct4, Sox2, Nanog and Klf4] are represented in green, while [Lin28A, Rex1, TRA-1–60 and TRA-1–81] are represented in red. DAPI marks nuclei (blue). One representative colony from each reprogrammed sample in reference to H1 and IMR90-1 cells is illustrated in this figure. HC (C7), ID-PD (C2), A53T-PD1 (C1) and A53T-PD2 (C2).