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Modulation of signalling pathways

JC Jesica R. Canizo
AR Amada E. Ynsaurralde Rivolta
CE Camila Vazquez Echegaray
MS Mariana Suvá
VA Virgilia Alberio
JA Juan F. Aller
AG Alejandra S. Guberman
DS Daniel F. Salamone
RA Ricardo H. Alberio
RA Ramiro Alberio
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MEK inhibition dose response curve: from day 5, embryos were cultured in N2B27 with DMSO (control) or in N2B27 with 0.4–10 μM of PD0325901 (TOCRIS, catalogue number 4192). Development was evaluated at day 7 and 8 and embryos were classified as BL: blastocyst, EB: expanded blastocyst, BHing: hatching blastocyst and BHed: hatched blastocyst. Hatched embryos were fixed with 4% of PFA at day 8 prior to immunostaining analysis. For MEK and Wnt inhibition experiments day 5 embryos were cultured in N2B27 with i) 10 μM of PD0325901, ii) 2.5 μM of IWP2 (Stemgent, catalogue number 04–0034), and iii) the combination of 10 μM PD0325901 and 2.5 μM IWP2. For t2iGö + LIF, t2iGö and LIF, a modified concentration of 1 μM CHIR (TOCRIS, catalogue number 4423), 1 μM of PD0325901, 2 μM of PKC inhibitor Gö6983 (AXON MEDCHEM,) and 20 ng/ml of h-LIF (Peprotech, catalogue number 300–05) were used.

Copyright and License information: The Author(s). ©2019
Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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