Figure 5.

Suppression of Nudt7 increases β-catenin signaling. (A) Canonical pathway was analyzed using IPA software (Qiagen, Redwood, CA, USA) from microarray data of Nudt7^+/+ and Nudt7^−/− colons treated with AOM/DSS. (B) Active z-score of disease and biological functions or molecular and cellular functions were analyzed using IPA software. (C) GSEA analysis using GEO datasets (CRC patient biopsy dataset, GSE41258 and Kras^G12D mutation CRC cell line dataset, GSE97023) and expression levels of Nudt7 and Ctnnb1. (D) Immunohistochemical staining with β-catenin and CK19 and positive cell count in Nudt7^+/+ and Nudt7^−/− mice treated with AOM/DSS (n = 4; 100× magnification; scale bars: 200 μm) and Kras^G12D-derived CRC patient (n = 4; β-catenin, 100× magnification, scale bars: 200 μm; CK19, 200× magnification, scale bars: 100 μm) and positive cells were counted for every 100 cells in 3 different fields. CK19 was used as a marker for epithelial tissue. Results are representative of at least 3 independent experiments. The dotted line boxes were enlarged in the bottom left corner of each image. (E) Transcriptional levels of β-catenin target genes and presented as the fold change compared with Nudt7^−/− mice. Rn18s was used as an endogenous control. Results are representative of at least 3 independent experiments. Values are means + SD. An unpaired Student’s t-test was used for statistical analysis. * p ≤ 0.05, ** p < 0.01, *** p < 0.01, **** p < 0.0001.