Fig. 5

WNT4 expression is decreased in the radiation-injured human and mouse intestinal tissues. A HE staining of the small intestines of normal control mouse (Normal) and mouse with 10.5 Gy of abdominal irradiation after 4 weeks (IR). Scale bars, 100 μm. B HE staining of the radiation-injured lesion (L) and paired adjacent non-injured intestinal tissue (N) in RE patient. Scale bars, 100 μm. C IF staining was performed using anti-lysozyme (labeled Paneth cells, red) and anti-EphB3 (green) antibodies in the small intestinal tissues of normal and IR mice. Nuclei were counterstained with DAPI (blue). Scale bars, 100 μm. D ISH analysis of Wnt4 expression in the small intestinal tissues of normal and IR mice. The crypt regions (red boxes) were magnified in the middle column, and the villous regions (black boxes) were magnified in the right column. Scale bars, 200 μm (left column) and 50 μm (middle and right columns). E The intensity of Wnt4 expression in the crypts and villus mesenchyme in the normal (n = 6) and IR (n = 6) mice was quantified using ImageJ software based on the ISH analysis in D. F RT-qPCR analysis of WNT4 expression in seven clinical samples of RE lesions (L) and paired adjacent non-injured intestinal tissues (N). G Correlation analysis between the relative WNT4 mRNA level and EphB3 mRNA level in the intestinal lesions of patients with RE (n = 18). Data are means ± SD. Statistical analysis was performed by unpaired Student’s t-test (E), paired Student’s t-test (F), and Pearson correlation coefficient test (G). ns, not significant, *p < 0.05, **p < 0.01