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Additional Supporting Information may be found in the online version of this article.

FilenameFormatSizeDescription
STEM_497_sm_SuppFig1.tif622KSupporting Information Figure 1. β-catenin-/- ESC are defective for endoderm/mesoderm differentiation. (A) qPCR expression analysis of endoderm markers (Sox17, and Gata6), mesoderm markers (Brachyury, and Eomes), and ectoderm marker (Nestin) upon embryoid body differentiation in wt and β-catenin-/- ESC.
STEM_497_sm_SuppFig2.tif1033KSupporting Information Figure 2. β-catenin-/- ESC exhibit decreased transcript levels of the pluripotency factors Oct4 and Nanog upon RA differentiation. (A) qRT-PCR analysis of Oct4, Nanog, Lrh-1, and β-catenin expression in wt and β-catenin-/- ESC upon RA-mediated differentiation. *, P < 0.05
STEM_497_sm_SuppFig3.tif827KSupporting Information Figure 3. CHIR99021 induces Lrh-1 in a β-catenin-dependent manner. (A) qPCR analysis of Lrh-1 Expression in wt and β-catenin-/- ESC upon 12 hours treatment with CHIR99021 (10 nM-5 μM) in the absence of LIF.
STEM_497_sm_SuppFig4.tif1055KSupporting Information Figure 4. Lrh-1-/- ESC exhibit decreased transcript levels of the pluripotency factors Oct4 and Nanog upon RA differentiation. (A) qRT-PCR analysis of Oct4, Nanog, Lrh-1, and β-catenin expression in wt and Lrh-1-/- ESC upon RA-mediated differentiation. *, P < 0.05
STEM_497_sm_SuppFig5.tif616KSupporting Information Figure 5. The induction of endoderm/mesoderm/ectoderm factors upon embryoid body differentiation is intact in Lrh-1-/- ESC. (A) qPCR expression analysis of endoderm markers (Sox17, and Gata6), mesoderm markers (Brachyury, and Eomes), and ectoderm marker (Nestin) upon embryoid body differentiation in wt and Lrh-1-/- ESC.
STEM_497_sm_SuppFig6.tif909KSupporting Information Figure 6. BIO is unable to maintain Oct4 expression in long-term culture. (A) Wild-type mES cells were cultured in the absence of LIF with media supplemented with BIO or DMSO vehicle for 7 days. Oct4 expression was examined by Western blot analysis and β-actin was probed as a loading control.
STEM_497_sm_SuppFig7.tif1362KSupporting Information Figure 7. Generation of β-catenin-/-(mycLrh-1) stably expressed ESC. (A) Western blot analysis of isolated clones. Heterologous recombinant colonies were cultured in the presence of LIF and G418. Transgene expression was confirmed by blotting for the myc-epitope alongside Lrh-1. All clones are absent for β-catenin, and β-actin served as a loading control.
STEM_497_sm_SuppTables1-3.doc74KSupporting Information Tables 1 to 3.

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