Figure 1: Evi/Wls is highly expressed in colon tumours.

(a,b) Immunohistochemical staining of normal or carcinoma tissue sections from colon, using the indicated antibodies. Images are representative of five cases. (a) In normal tissue, Evi/Wls expression is mostly undetectable, whereas Wnt3 is weakly expressed in most epithelial cells but more highly expressed in the Paneth-like/endocrine cells, which have a granular appearance (arrow). Keratin20 staining marks the top of the intestinal crypts and Eph receptor B2 staining the bottom. (b) In matched colon carcinomas, Evi/Wls and Wnt3 are highly expressed. (a,b) Scale bars, 60 μm, Wnt3 (enlarged) 20 μm. (c) Evi/Wls and Wnt3 are overexpressed in colon cancer. Results of immunostainings for Evi/Wls and Wnt3 on a colon cancer tissue array. Scores provided independently by three different investigators are shown as average. Staining intensity: 0=no staining, 1=weak staining, 2=moderate staining, 3=strong staining; percentage bin of cells: 0=<10%, 1=10–25%, 2=25–50%, 3=50–75%, 4=>75%). Grey scale represents the number of tumours. (d) Wnt3 is overexpressed in adenocarcinomas. Microarray-based expression data (Oncomine database, TCGA data set) were analysed for the expression of Wnt3 in normal colon(19)/rectum(3) versus adenocarcinomas (101). Preprocessed data from the array is log²-transformed and median-centred. The box plots show medians as line within the box, 25th and 75th percentiles as sides of the box, 10th and 90th percentiles as error bars, and outliers as circles. Significance of the differences in expression was calculated using the Student’s t-test. The specificity of the Wnt3 antibody was tested by knockdown experiments (Supplementary Fig. S1a). Evi/Wls antibody is described in Augustin et al.¹⁰