1. The Intestinal Wnt/TCF Signature
Laurens G. Van der Flier, Jacob Sabates–Bellver, Irma Oving, Andrea Haegebarth, Mariagrazia De Palo, Marcello Anti, Marielle E. Van Gijn, Saskia Suijkerbuijk, Marc Van de Wetering, Giancarlo Marra, Hans Clevers 
Gastroenterology 
Volume 132, Issue 2, Pages (February 2007)
DOI: /j.gastro
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2. Figure 1
Over expression of dnTCFs inhibits the Wnt pathway in CRC cells. TCF/β-catenin driven transcription is abrogated by over expression of dnTCF1 or dnTCF4 in (A) LS174T cells and (B) DLD1 cells. The activity of the TCF reporter, pTopGlow (□, TOP), and the control, pFopGlow (■, FOP), after 20 hours with or without doxycycline treatment is shown. Parental cells that expressed the tetracycline repressor (TR) were used as controls. Renilla luciferase levels were used as transfection controls. (C) Venn diagram showing the (overlapping) down-regulated probes on dnTCF1 or dnTCF4 induction. Selection is based on down-regulation in 2 or more of the 4 cell-line transfectants used in this study. Not shown in the figure is the overlap in 6 unique probes between LS174T/dnTCF1 and DLD1/dnTCF4 and 6 probes between LS174T/dnTCF4 and DLD1/dnTCF1.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2007 AGA Institute Terms and Conditions

3. Figure 2
The intestinal Wnt/TCF signature gene set. TCF/β-catenin target genes were selected by comparing the 387 probes (lower circle) obtained with the procedure described in Figure 1C with the 8307 probes whose expression levels in human tumors were increased significantly, relative to those in normal mucosa. The left upper circle represents 3319 probes increased in adenomas, whereas the right upper circle represents 4988 probes increased in carcinomas (1971 were increased in both tumor types). With this approach, we classified 51 genes (71 probes) up-regulated in adenomas, 36 genes (40 probes) up-regulated in carcinomas, and 121 genes (144 probes) up-regulated in both tumor types as β-catenin/TCF target genes. These gene lists are reported in Supplementary Table 1 (supplementary material online at
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2007 AGA Institute Terms and Conditions

4. Figure 3
Confirmation and classification of TCF target genes by in situ hybridization on intestines of adult APCmin mice. All Wnt signature genes tested were expressed in (A, C, E, and G) wild type crypts and in (B, D, F, and H) adenomas of Apcmin mice. The staining patterns in wt crypts could be divided into 3 categories. (A and B) An example of the expression of a gene within the first category, the mouse ortholog of HSPC111, is shown. In addition to expression within the proliferative compartment of crypts and adenomas, a decreasing gradient of signal is observed along the base of the villus. (C and D) Another gene in this category, the mouse ortholog for NOL1, showed expression restricted to the proliferative compartment of crypts and adenomas, but lacked significant expression along the villus. (E and F) The second staining pattern involves the Paneth cells at the bottom of the crypts, as represented by the mouse ortholog for MMP7. (G and H) In situ hybridizations for the mouse ortholog of ASCL2 defined a third category that revealed expression in adenomas and in a few cells near the crypt bottom. This position coincides with the location of the elusive crypt stem cells. Size bars represent 50 μm, and adenomas are encircled by dotted lines.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2007 AGA Institute Terms and Conditions