Vacik T , Stubbs JL , Lemke G .

EY017478 NEI NIH HHS , R01 EY017478 NEI NIH HHS

	Figure 6. DnTcf7l2 is essential for Xenopus forebrain development. (A) A 59 RACE experiment in X. laevis reveals the existence of an alternative XTcf7l2 first exon in intron 5, which contains an ATG start codon in-frame with Ser 180 in XTcf7l2 exon 6. The positions of the translation-blocking (MO-ATG) and splicing- blocking (MO-SPL) morpholinos are indicated. Arrows represent the primers used for qRTCR. (B) In situ hybridization analysis of dnTcf7l2 expression at stages 13, 15, and 28 of X. laevis development. (A) Anterior; (P) posterior; (D) dorsal; (V) ventral; (bp) blastopore. (C) Knockdown of XdnTcf7l2 with MO-ATG or MO-SPL leads to embryos with severely truncated anterior head regions at stage 40. Phenotype ranges from moderate (gray bars), showing small head and small eyes, to strong (black bars), where no head and no eyes are present.
	Figure S8. Expression of Spemann organizer and anterior neural markers during embryogenesis of Xenopus morphants lacking XdnTcf7l2. In situ hybridization was carried out for: the organizer marker chordin (Chd) at st. 10.5 (1st column, vegetal view), the organizer+anterior neuroectodermal marker Otx2 (Otx2) at st. 12.5 (2nd column, anterior up); the anterior neuroectodermal marker Otx2 at st. 20 (3rd column, anterior on); and the midbrain/hindbrain boundary marker engailed 2 (En2) at st. 20 (4th column, anterior on). These hybridizations were carried out in embryos injected with morpholinos in one animal blastomere (left, injected in all panels) of 2-cell stage embryos, with the uninjected side (right in all panels) as a control. (Injected side marked with lacZ.) Morpholinos used were: control (top row); ATG morpholino (middle row), and splicing morpholino (bottom row). Percent tally of results from repeated analyses in multiple embryos (n = 18-46, indicated by white numbers), with respect to level of marker expression on the injected side (same, reduced, or absent), is given in the histograms below each column. Anterior neural development is unaffected by either morpholino prior to st. 12.5, the time at which XdnTcf7l2 mRNA first appears.
	Figure S9. Misexpression of XdnTcf7l2 in Xenopus laevis embryos results in multiple developmental anomalies. (A) As in the mouse, Tcf7l2 and dnTcf7l2 in Xenopus exist in multiple isoforms due to alternative splicing of exons 13-16. We identified the indicated XdnTcf7l2 isoforms, designated 1-4, as most abundant in stage 13 and 24 embryos, with their relative abundance at each stage indicated as a percentage. Color coding of XdnTcf7l2 protein domains within exons is as for Figure 3. X indicates the presence of a stop codon. Isoforms 1-3 contain the short SFLSS motif of Pukrop et al. (2001) in exon 9 (indicated by , while isoform 4 does not (indicated by . (B) We injected mRNAs encoding the indicated isoforms, either singly or in combination (1+2 and 3+4), into either the two dorsal blastomeres (orsal injections or one ventral blastomere (entral injections of 4-cell-stage embryos, with nuclear -galactosidase mRNA co-injected as a marker (or control). Embryos were scored for the indicated patterning phenotypes at st. 30-32. N indicates number of embryos scored. (C) Selected examples of patterning defects observed. Although these studies indicate that XdnTcf7l2 has potent developmental effects in vivo, there are two important caveats to interpreting these effects in a biological context. First, we are over-expressing exogenous XdnTcf7l2 much earlier in development than its normal first appearance at st. 13, and therefore may be affecting developmental events in which XdnTcf7l2 does not participate. And second, it is possible that this early expression of XdnTcf7l2 interferes with (inhibits) the activity of other XTcf proteins that may act earlier in development than does XTcf7l2.

Angus-Hill, T-cell factor 4 functions as a tumor suppressor whose disruption modulates colon cell proliferation and tumorigenesis. 2011, Pubmed

Angus-Hill, T-cell factor 4 functions as a tumor suppressor whose disruption modulates colon cell proliferation and tumorigenesis. 2011, Pubmed
Arce, Diversity of LEF/TCF action in development and disease. 2006, Pubmed
Barbieri, A homeobox gene, vax2, controls the patterning of the eye dorsoventral axis. 1999, Pubmed , Xenbase
Bertuzzi, The homeodomain protein vax1 is required for axon guidance and major tract formation in the developing forebrain. 1999, Pubmed
Bharti, Lack of the ventral anterior homeodomain transcription factor VAX1 leads to induction of a second pituitary. 2011, Pubmed
Brinkmeier, TCF4 deficiency expands ventral diencephalon signaling and increases induction of pituitary progenitors. 2007, Pubmed
Capdevila, Patterning mechanisms controlling vertebrate limb development. 2001, Pubmed
Cho, TCF-4 binds beta-catenin and is expressed in distinct regions of the embryonic brain and limbs. 1998, Pubmed , Xenbase
de Lau, LEF1 turns over a new leaf. 2001, Pubmed
Echelard, Sonic hedgehog, a member of a family of putative signaling molecules, is implicated in the regulation of CNS polarity. 1993, Pubmed
Fujimura, Spatial and temporal regulation of Wnt/beta-catenin signaling is essential for development of the retinal pigment epithelium. 2009, Pubmed
Grove, Wnt signaling meets internal dissent. 2011, Pubmed , Xenbase
Hallonet, Vax1, a novel homeobox-containing gene, directs development of the basal forebrain and visual system. 1999, Pubmed , Xenbase
Hoppler, Wnt signalling: variety at the core. 2007, Pubmed
Hovanes, Beta-catenin-sensitive isoforms of lymphoid enhancer factor-1 are selectively expressed in colon cancer. 2001, Pubmed
Hurlstone, T-cell factors: turn-ons and turn-offs. 2002, Pubmed
Jessell, Neuronal specification in the spinal cord: inductive signals and transcriptional codes. 2000, Pubmed
Ji, An integrated software system for analyzing ChIP-chip and ChIP-seq data. 2008, Pubmed
Kao, The entire mesodermal mantle behaves as Spemann's organizer in dorsoanterior enhanced Xenopus laevis embryos. 1988, Pubmed , Xenbase
Kim, Hedgehog-regulated localization of Vax2 controls eye development. 2006, Pubmed
Kim, Repressor activity of Headless/Tcf3 is essential for vertebrate head formation. 2000, Pubmed
Korinek, Constitutive transcriptional activation by a beta-catenin-Tcf complex in APC-/- colon carcinoma. 1997, Pubmed
Korinek, Depletion of epithelial stem-cell compartments in the small intestine of mice lacking Tcf-4. 1998, Pubmed
Korinek, Two members of the Tcf family implicated in Wnt/beta-catenin signaling during embryogenesis in the mouse. 1998, Pubmed
Kunz, Autoregulation of canonical Wnt signaling controls midbrain development. 2004, Pubmed , Xenbase
Lee, Chromatin immunoprecipitation and microarray-based analysis of protein location. 2006, Pubmed
Liu, A role for Xvax2 in controlling proliferation of Xenopus ventral eye and brain progenitors. 2008, Pubmed , Xenbase
Logan, The Wnt signaling pathway in development and disease. 2004, Pubmed
MacDonald, Wnt/beta-catenin signaling: components, mechanisms, and diseases. 2009, Pubmed , Xenbase
Maretto, Mapping Wnt/beta-catenin signaling during mouse development and in colorectal tumors. 2003, Pubmed
Mayor, VISTA : visualizing global DNA sequence alignments of arbitrary length. 2000, Pubmed
Mui, Vax genes ventralize the embryonic eye. 2005, Pubmed
Mui, The homeodomain protein Vax2 patterns the dorsoventral and nasotemporal axes of the eye. 2002, Pubmed
Mukhopadhyay, Dickkopf1 is required for embryonic head induction and limb morphogenesis in the mouse. 2001, Pubmed , Xenbase
Paridaen, Apc1-mediated antagonism of Wnt/beta-catenin signaling is required for retino-tectal pathfinding in the zebrafish. 2009, Pubmed
Parr, Mouse Wnt genes exhibit discrete domains of expression in the early embryonic CNS and limb buds. 1993, Pubmed
Petersen, Wnt signaling and the polarity of the primary body axis. 2009, Pubmed
Schulte, Misexpression of the Emx-related homeobox genes cVax and mVax2 ventralizes the retina and perturbs the retinotectal map. 1999, Pubmed
Sharov, CisView: a browser and database of cis-regulatory modules predicted in the mouse genome. 2006, Pubmed
Sousa, Sonic hedgehog functions through dynamic changes in temporal competence in the developing forebrain. 2010, Pubmed
Struewing, The balance of TCF7L2 variants with differential activities in Wnt-signaling is regulated by lithium in a GSK3beta-independent manner. 2010, Pubmed
Stubbs, The forkhead protein Foxj1 specifies node-like cilia in Xenopus and zebrafish embryos. 2008, Pubmed , Xenbase
Take-uchi, Hedgehog signalling maintains the optic stalk-retinal interface through the regulation of Vax gene activity. 2003, Pubmed , Xenbase
Ulloa, Wnt won the war: antagonistic role of Wnt over Shh controls dorso-ventral patterning of the vertebrate neural tube. 2010, Pubmed
Van de Wetering, Extensive alternative splicing and dual promoter usage generate Tcf-1 protein isoforms with differential transcription control properties. 1996, Pubmed
Veien, Canonical Wnt signaling is required for the maintenance of dorsal retinal identity. 2008, Pubmed
Weise, Alternative splicing of Tcf7l2 transcripts generates protein variants with differential promoter-binding and transcriptional activation properties at Wnt/beta-catenin targets. 2010, Pubmed
Wiley, GSK-3beta and alpha-catenin binding regions of beta-catenin exert opposing effects on the terminal ventral optic axonal projection. 2008, Pubmed , Xenbase
Zhou, Ocular coloboma and dorsoventral neuroretinal patterning defects in Lrp6 mutant eyes. 2008, Pubmed