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. 2017 Nov 20;7:15852. doi: 10.1038/s41598-017-15626-9

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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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C-Myc overexpression promotes chicken cell proliferation and miR-17-92 cluster expression. (a) Effect of c-Myc overexpression on the proliferation of DF1 cells. Cells were transfected with either pCMV-HA-c-Myc or pCMV-HA vector, and cell proliferation was assayed at the designated time points after transfection using the CCK-8 kit. (b) qRT-PCR analysis of PCNA, Cyclin D1 and Ki67 in DF1 cells. Cells were transfected with pCMV-HA-c-Myc or pCMV-HA vector at the designated time points after transfection, and total RNA was isolated and gene expression was assessed using qRT-PCR. Gene expression was normalized to NONO mRNA level. Fold change is relative to pCMV-HA vector at 24 h after transfection. (c) DF1 cells were transfected with either pCMV-HA-c-Myc or pCMV-HA vector at the designated time points after transfection, total RNA was isolated and the expression of pri-miR-17-92 and its mature miRNA members was quantified using qRT-PCR and stem-loop qRT-PCR, respectively. The pri-miR-17 -92 expression was normalized to NONO mRNA level, and the mature miRNA expression was normalized to U6 snRNA expression level. Fold change is relative to pCMV-HA vector at 24 h after transfection. All data are representative of three independent experiments and shown as the mean ± SEM. *p < 0.05; **p < 0.01; determined by two-tailed Student’s t-test.