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. 2015 Apr 15;142(8):1528–1541. doi: 10.1242/dev.119271

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© 2015. Published by The Company of Biologists Ltd

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.

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Fig. 3. — Characterisation of HPSC-derived epicardium. (A) Analysis of epicardial markers in H9 and BHX-derived epicardium-like cells by qRT-PCR. Significant differences compared with H9 are indicated in black, whereas intergroup differences are in red. ***P<0.001, **P<0.01, *P<0.05. (B) Western blot showing WT1 and TCF21 expression in H9 and H9-derived day 10 (EPI D10) epicardial cells. β-Actin was used as a loading control. (C) Immunocytochemistry for WT1 and BNC1 in EPI D10. Rabbit IgG isotype used as negative control. Scale bars: 100 μm. (D) (a) Flow cytometry analysis of WT1⁺, TCF21⁺ and WT1⁺/TCF21⁺ cells in EPI D10. Rabbit IgG isotype was used as a negative control. (b) Percentage of cells uniquely expressing WT1, TCF21 and WT1/TCF21, and cells not expressing either of the two markers. (E) (a) Expression of epicardial markers in EPI D10 and human foetal epicardial outgrowths (FOETAL EPI) by qRT-PCR. Significant differences compared with LM are in black, whereas intergroup differences are in red. ***P<0.001, **P<0.01, *P<0.05. (b) Bright-field images of EPI D10 and FOETAL EPI. Scale bars: 100 μm.