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. 2013 Jun 27;14(7):13482–13496. doi: 10.3390/ijms140713482

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© 2013 by the authors; licensee MDPI, Basel, Switzerland

This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/3.0/).

PMC Copyright notice

The effects of miR-125b overexpression on TGFβ signaling are mediated by the BMP4 co-receptor, Dies1. (A) qPCR analysis of the expression levels of BMP4 (Id1, Id3) and Nodal/Activin (Nodal, Cripto, Lefty1, Lefty2) target genes upon miR-125b overexpression; (B) Analysis of the phenotype of ESCs co-transfected with the indicated pre-miR and with the vector expressing Dies1 lacking its 3′UTR or with the empty vector (mock). The expression of stemness (Oct3/4) and neuroectodermal (Sox1) markers was analyzed by immunostaining in cells differentiated as SFEBs for four days. Scale bar: 20 μm; (C) q-PCR analysis of the effects of Dies1 re-expression in ESCs transfected with the indicated pre-miR. After four days of differentiation, the expression of stemness (Oct3/4, Nanog) and epiblast (Fgf5) markers was analyzed; (D) q-PCR analysis of the expression of BMP4 (Id1) and Nodal/Activin (Lefty1 and Lefty2) targets in four-day differentiated ESCs re-expressing, or not, Dies1 upon miR-125b overexpression. Data in (C) and (D) are shown as fold changes relative to cognate controls; (E) The effects of miR-125a and b overexpression on Lin28 level were analyzed in four-day differentiated SFEBs by means of q-PCR (left panel) and Western blot (right panel). Data are expressed as fold change relative to the control (*p < 0.05).

The effects of miR-125b overexpression on TGFβ signaling are mediated by the BMP4 co-receptor, Dies1. (A) qPCR analysis of the expression levels of BMP4 (Id1, Id3) and Nodal/Activin (Nodal, Cripto, Lefty1, Lefty2) target genes upon miR-125b overexpression; (B) Analysis of the phenotype of ESCs co-transfected with the indicated pre-miR and with the vector expressing Dies1 lacking its 3′UTR or with the empty vector (mock). The expression of stemness (Oct3/4) and neuroectodermal (Sox1) markers was analyzed by immunostaining in cells differentiated as SFEBs for four days. Scale bar: 20 μm; (C) q-PCR analysis of the effects of Dies1 re-expression in ESCs transfected with the indicated pre-miR. After four days of differentiation, the expression of stemness (Oct3/4, Nanog) and epiblast (Fgf5) markers was analyzed; (D) q-PCR analysis of the expression of BMP4 (Id1) and Nodal/Activin (Lefty1 and Lefty2) targets in four-day differentiated ESCs re-expressing, or not, Dies1 upon miR-125b overexpression. Data in (C) and (D) are shown as fold changes relative to cognate controls; (E) The effects of miR-125a and b overexpression on Lin28 level were analyzed in four-day differentiated SFEBs by means of q-PCR (left panel) and Western blot (right panel). Data are expressed as fold change relative to the control (*p < 0.05).