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Fig. 1. Isl1 positively regulates Dkk1 expression. A, B. Isl1OE ES line at day 4 of cardiac differentiation (A) and Isl1-overexpressing endodermal NFPE cells (B) show increased Dkk1 expression as determined by real-time RT-PCR. Dkk1 gene expression levels were normalized to Hprt (A) or Gapdh (B); n = 3. C. Schematic representation of the 1.3 kb promoter region upstream of the Dkk1 transcription site containing two predicted Isl1-binding sites. Deletion constructs generated are given. D. Bar graph illustrating the luciferase activity. NFPE cells were transiently transfected with a pGL3-basic luciferase reporter or with the same vector encompassing the 1.3 kb Dkk1 promoter fragment, together with an expression vector containing GFP control or Isl1. In addition, modified pGL3-basic-Dkk1-promoter constructs were used, in which either one or both Isl1-binding sites were deleted. The luciferase levels were normalized for the β-galactosidase activity of a co-transfected RSV-βGal and shown as luciferase activity relative to pGL3-basic plus GFP control (RLU, relative light units). n = 6–8. For all panels: Mean values with standard errors are given. *p < 0.05, **p < 0.01 and ***p < 0.001 with Student’ T-test. Image published in: Guo Y et al. (2019) © 2019 The Authors. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivatives license Larger Image Printer Friendly View

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