Figure 3

Functional phenotype in WNT1-injected mice. a Intestinal section stained with antibodies to β-galactosidase (β-gal; red) and green fluorescent protein (GFP; green), and counterstained with Hoechst dye (blue) in a WNT1-injected Wnt-reporter mouse. Arrowheads denote GFP-expressing WNT1-infected cells. Higher magnification of white box in inset. Dashed line represents epithelial-mesenchymal boundary. b Quantitation of intestinal Wnt-receiving cells in the proximal, mid, and distal small intestine (PSI, MSI, and DSI) by immunostaining with antibodies to β-gal. The number of crypt-villus units containing Wnt-receiving cells was counted in DsRed-injected (blue bars) and WNT1-injected (red bars) Wnt-reporter mice and represented as a ratio to total crypt-villus units counted (n = 3 for all three regions; MSI, p = 0.0079). c Quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) assay for LacZ expression in isolated intestinal crypt, villus, and mesenchymal cells from WNT1- and DsRed-infected Wnt-reporter mice. Graph displays triplicate samples from representative animals. Data displayed as mean ± std. d qRT-PCR for gene expression of canonical Wnt target genes: c-Myc, EphrinB1, EphB2, and EphB3. All qRT-PCR samples normalized to internal reference gene, represented relative to DsRed-infected cells and run in triplicate. e Intestinal crypt section from WNT1-injected mouse stained with antibodies to Ki67 (red) and GFP (green). Solid white bar indicates crypt-villus junction. Arrowheads denote Ki67-positive epithelial cells outside of normal proliferative zone. Dashed line indicates epithelial-mesenchymal boundary. f Polyp from PSI of WNT1-infected mouse stained with antibodies to Ki67 (red) and Hoechst (blue). Bar = 25 μm. g Quantitation of intestinal tumors from DsRed- and WNT1-injected mice.