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. 2018 Aug 28;8:12977. doi: 10.1038/s41598-018-30939-z

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© The Author(s) 2018

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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ATC application downregulated pluripotency markers in hESCs. (A) Bright field images of single ESCs and integrin-bound MBs (arrows). Confocal fluorescence images of single hESCs stained with DAPI (blue), Oct4 (magenta), Nanog (yellow), and Sox2 (green) with and without ATC application (30 min). Scale bars: 50 µm. (B) Percentages of hESCs with Oct4, Nanog, and Sox2 after ATC treatment compared with control. (C) Bright field images of a hESC colony and integrin-bound MBs (arrows). Confocal fluorescence images showing hESCs stained with DAPI (blue), Oct4 (magenta), Nanog (yellow), and Sox2 (green) with and without ATC application. Scale bars: 50 µm. (D) Corrected fluorescence intensity measurement of hESCs with Nanog, Oct4, and Sox2 after ATC treatment. (E) Immunoblot analysis of protein expression in hESCs. All quantifications were from at least 3 independent experiments with two replicates per experiment. Unpaired t test p values * < 0.05, **<0.01, and ***<0.001; n.s, not significant.