File:Ectoderm Specification of Human Embryonic Stem Cells.png

Ectoderm Specification of Human Embryonic Stem Cells

(A) Schematic of the protocol used to induce ectoderm specification of hESCs in the presence of an inhibitor of a ϒ-secretase inhibitor (DAPT) or vehicle control (ethanol). (B) Morphological analysis of phase images of hESCs colonies at different timepoints throughout the ectoderm specification protocol. DAPT-treatment accelerates colony differentiation as made evident by a qualitative increase in number of cells displaying a flattened morphological appearance. (C) Undifferentiated hESCs are negative for P63 (red) and AP2α (green), and are positive for the pluripotency markers SOX2 (red) and OCT4 (green). As cells differentiate, DAPT-treated colonies display an increase in P63 and AP2α expression and a loss in OCT4 and SOX2 at 3 and 6 days in differentiation conditions. Dotted line outlines colony edge and the asterisk marks undifferentiated areas of the colonies. (D) Quantitative real-time PCR (qRT-PCR) analysis of mRNA levels of POU5F1 during differentiation as compared to undifferentiated hESCs (n = 6 independent differentiation experiments for each bar). (E) qRT-PCR analysis of mRNA levels of TFAP2A during differentiation as compared to undifferentiated hESCs (n = 3 independent differentiation experiments for each bar). All data are ± SEM (** 0.001<p<0.01, *0.01<p<0.05). Scale bars represent 100μm.

z5019306

http://dx.doi.org/10.1371/journal.pone.0122493.g001

References

<pubmed>25849374</pubmed>

Copyright

© 2015 Tadeu et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited

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