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. 2017 May 4;7:1500. doi: 10.1038/s41598-017-01628-0

Figure 1.

Figure 1

Cnot3 is required for cardiac differentiation. (a) NT shRNA-(shNT) or Cnot3 shRNA (shCnot3)-transduced human ESCs were induced to differentiation (see Methods for details) for 12 days, and the amount of beating colonies were counted and plotted as mean ± SEM from three independent experiments. (b) The expression of cardiac specific gene was determined by qPCR 12 days after hESCs differentiation. β-actin was used as endogenous control and values were plotted as mean ± SEM from three independent experiments. (c) GFP or Cnot3-overexpressing human ESCs were induced to differentiation (see Methods for details) for 12 days, and the amount of beating colonies were counted and plotted as mean ± SEM from three independent experiments. (d) The expression of myocyte specific gene was determined by qPCR 12 days after hESCs differentiation. β-actin was used as endogenous control and values were plotted as mean ± SEM from three independent experiments. (e,f) Inducible-shCnot3 or Inducible-shNT hESCs were induced to differentiation, and doxycycline was added during the differentiation process at the indicated time points. The effects of Cnot3 depletion on myocyte specific gene expression at different stages were determined by qPCR. β-actin was used as endogenous control and values were plotted as mean ± SEM from three independent experiments. (g,h) Inducible-Cnot3 or Inducible-GFP overexpression hESCs were induced to differentiation, and Doxycycline was added during the differentiation at the indicated time points. Effects of Cnot3 depletion at different stage on myocyte specific gene expression were determined by RT-qPCR. β-actin was used as endogenous control and values were plotted as mean ± SEM from three independent experiments.