FGF-signalling in the differentiation of mouse ES cells towards ...

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Figure 2: Activation of FGFRc-isoforms boosts mesendoderm but inhibits DE marker expression. Using GFPreportercell lines T-GFP, Gsc-GFP and Sox17-GFP, we differentiated cells for 3 (T-GFP cell line only) and 5 daysin BMP4- or activin-containing media, adding different FGFs. Cells were analysed by a FACS. A) Table of FGF –FGFR binding of selected FGFs used in this paper. Modified from {Olsen, 2006 #25;Mason, 2007 #21;Ornitz, 1996#20;Zhang, 2006 #19}. B-C) Cells were differentiated in 10 ng/ml BMP4 (= mesoderm-induction) or 1 ng/mlactivin (= posterior streak/ mesoderm-induction) w/wo FGFs, and expression of T-GFP was measured at days 3 and5. D) Gsc-GFP cells were differentiated in 30 ng/ml activin w/wo FGFs, and expression of GFP was measured atday 5. E) Sox17-GFP cells were differentiated in 30 ng/ml activin w/wo FGFs, and expression of GFP wasmeasured at day 5. The mean expression ± S.E.M. of 3 independent experiments is shown, using a Student’s t-testfor the statistical analysis: * = P < 0,05; ** = P < 0,01 compared to the BMP4 or activin conditions.76
Figure 3: Activation of FGFRb or FGFRc-isoforms differentially affects the expression of PS, DE andmesoderm markers. Sox17-GFP cells were differentiated in media containing BMP4 or activin w/wo FGFs for 3-5days before harvest. Cells were stained for markers of PS, DE or mesoderm and analysed using a FACS. A & B)Cells were stained for T and analysed on days 3 and 5. Shown here are the relative number of T + cells in the Sox17-GFP Hi fraction A), or the Sox17-GFP –/Lo fraction B). The mean expression ± S.E.M. of 2 independent experimentsare shown, no statistical analysis performed. C-G) Cells were stained for FLK1 and EpCAM and analysed in eithersingle channel for C) Sox17-GFP, D) FLK1, E) EpCAM; or in multichannel for the Sox17-GFP Hi fraction F), orSox17-GFP –/Lo fraction G) dividing data into four fractions: FLK1 – /EpCAM + ; FLK1 + /EpCAM + ; FLK1 – /EpCAM – ;FLK1 + /EpCAM – . The mean expression ± S.E.M. of 3 independent experiments is shown, using a Student’s paired,two-tailed t-test for the statistical analysis: * = P < 0,05; ** = P < 0,01 compared to the activin conditions.77
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PhD thesisCand.scient. Janny Marie
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ResuméSukkersyge er en sygdom der
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Table of contents1
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ICMinner cell massIdInhibitor of di
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cell mass regenerates probably thro
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Figure 1-1: Early embryo developmen
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Figure 1-3: Regional expression of
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The pluripotent stateThe pluripoten
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There are four membrane-bound FGFRs
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2. AimsThe aim of this study was to
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Developmental Biology 330 (2009) 28
Page 32 and 33: 288 M. Hansson et al. / Development
Page 50 and 51: Figure S2Figure S2: A subpopulation
Page 52 and 53: Figure S4Figure S4: Expression of T
Page 54 and 55: Figure S6Figure S6: qRT-PCR analyse
Page 56 and 57: epithelium; Cdx2, expressed posteri
Page 58 and 59: Figure 4-4: A high FGF4-concentrati
Page 60 and 61: Figure 4-6: A 3-step protocol does
Page 63 and 64: 5. Paper IIFGFR(IIIc)-activation in
Page 65 and 66: AbstractProgress in embryonic stem
Page 67 and 68: variants in their Ig-like domain II
Page 69 and 70: Cell count and proliferation assayC
Page 71 and 72: influence on the numbers of Sox17-G
Page 73 and 74: undifferentiated cells, we found th
Page 75 and 76: FGF4, 5, FGF8b and FGFR1, are expre
Page 77 and 78: with EdU-stain (blue sample); and w
Page 79 and 80: Olsen, S.K., J.Y. Li, C. Bromleigh,
Page 81: FiguresFigure 1: Screen for FGFR-is
Page 87 and 88: Opposite, Figure 6: In the absence
Page 89 and 90: 6. General discussionEndoderm diffe
Page 91 and 92: Overall, the multitude of FGF-signa
Page 93 and 94: transplantation is the spread of an
Page 96: AcknowledgementsThe work presented
Page 99 and 100: Chambers, I., D. Colby, M. Robertso
Page 101 and 102: Hawkins, V.J. Wroblewski, D.S. Li,
Page 103 and 104: Nishikawa, S.I., S. Nishikawa, M. H
Page 105 and 106: Tanimizu, N., H. Saito, K. Mostov,
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