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. 2017 Dec 14;7:17572. doi: 10.1038/s41598-017-17828-7

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© The Author(s) 2017

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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Dual regulation of endocytic genes by the PRC2 complex and ESCC miRNAs. (a) RT-qPCR analysis of PRC2 target genes upon Ezh2 knockdown in V6.5 and R1 mESCs. mRNA expression is normalized to Gapdh and represented relative to scrambled shRNA. Scrambled shRNA is represented as a dashed line at 1. (b) Graph showing enrichment of SUZ12 at Cav1, Cdh2, Tgfbr1, Tgfbr2 and Tgfbr3 promoters by chromatin immunoprecipitation using IgG or SUZ12 specific antibody. (c) RT-qPCR analysis for endocytic gene expression in MEFs in the presence of exogenous miR-294, 12 and 24 hrs post-transfection. Mock is represented as a dashed line at 1. (d) RT-qPCR analysis of PRC2 target genes in Dgcr8 KO mESCs upon Ezh2 knockdown, and in the presence of exogenous miR-294. mRNA expression is normalized to Gapdh and represented relative to scrambled shRNA. Scrambled shRNA is represented as a dashed line at 1. (e) Luciferase analysis of Cav1 3′UTR, Cdh2 (N-Cad) ORF and human Tgfbr2 3′UTR. MEFs were co-transfected in the presence or absence of miR-294 and respective pSICHECK2 vectors. Renilla luciferase activity levels were normalized to firefly luciferase, which served as an internal control. Mock transfection level is represented as a dashed line at 1. For all experiments, error bars represent mean ± S.D for experiments in triplicates (N = 3). *p < 0.05; **p < 0.01; ***p < 0.001 by Students T-test.