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Stem Cells International
Volume 2017 (2017), Article ID 1769298, 10 pages
Research Article

miR-142-3p Contributes to Early Cardiac Fate Decision of Embryonic Stem Cells

1Key Laboratory of Stem Cell Biology and Laboratory of Molecular Cardiology, Institute of Health Sciences, Shanghai Jiao Tong University School of Medicine (SJTUSM) and Shanghai Institutes for Biological Sciences (SIBS), Chinese Academy of Sciences (CAS), Shanghai, China
2Department of Cardiology, Shanghai Xuhui District Central Hospital, Shanghai, China

Correspondence should be addressed to Huang-Tian Yang

Received 26 January 2017; Accepted 9 April 2017; Published 5 June 2017

Academic Editor: Yanfang Chen

Copyright © 2017 Zhong-Yan Chen et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Supplementary Material

Table S1. List of primer sequences for PCR detection. Figure S1. Isolation and characterization of the samples for microRNA microarray assay. (A) Schematic diagram of the strategy for sample collection. (B) FACS sorting of T-GFP- and T-GFP+ subpopulations and the purity detection. (C) FACS sorting of FLK1-/CXCR4- and FLK1+/CXCR4+ subpopulations and the purity detection. (D) RT-PCR analysis of T expression in unsorted, T-GFP- and T-GFP+ cells. (E) RT-PCR analysis of cardiac progenitor marker genes in unsorted, FLK1- /CXCR4- and FLK1+/CXCR4+ subpopulations. Figure S2. Knockdown of miR-142-3p does not affect the self-renewal and cardiomyocyte differentiation of ESCs. ESCs were transfected with 100 nM miR-142-3p inhibitor or scramble control for 48h. (A) qRT-PCR analysis of miR-142-3p in ESCs transfection with 100 nM miR-142-3p inhibitor or scramble control. (B) ALP staining of the colonies of ESCs (a-b). Immunostaining analysis of OCT4 (c-d) and NANOG (e-f). scale bar: a-b =100 mm,c-f =50 mm. (C) qRT-PCR analysis for the expression of the pluripotency marker genes. n=3. (D) Flow cytometry analysis of SSEA1. n=3. (E) The percentage of EBs with contracting clusters during differentiation. n=3. Figure S3. miR-142-3p does not directly target to the 3′UTR of Mesp1. (A) RNAHybrid predicts the binding of miR-142-3p to the 3′UTR of Mesp1. (B) Luciferase assay determined in HEK293T cells that were transfected with the 3′UTR reporter construct together with miR-142-3p mimics or scramble. n=3.

  1. Supplementary material