Fig. 6

AAV-mediated WNT4 supplementation inhibits EphB3 expression in small intestinal crypts in mice with radiation injury. The mice received 10.5 Gy of abdominal irradiation or sham-irradiation as control. The next day, AAV-vector or AAV-Wnt4 was intraperitoneally injected into the irradiated mice. At 4 weeks after irradiation, the small intestinal tissues were collected and analyzed. A The overexpression of WNT4 in the small intestines of irradiated mice treated with AAV-Wnt4 was confirmed by IF staining with anti-Flag antibody (green). Nuclei were counterstained with DAPI (blue). Scale bars, 100 μm. Dashed lines indicate crypt area. a: apical; b: basal. B RT-qPCR analysis of Wnt4 expression in the entire small intestinal tissues of the irradiated mice treated with AAV-vector (n = 5) or AAV-Wnt4 (n = 5) or the sham-irradiated control mice (n = 3). C IF staining was performed using anti-lysozyme (labeled Paneth cells, red) and anti-EphB3 (green) antibodies in the small intestinal tissues of the three groups of mice. Nuclei were counterstained with DAPI (blue). Scale bars, 100 μm. Dashed lines indicate crypt area. a: apical; b: basal. D The intensity of EphB3 expression and the height of the EphB3-positive area were quantified using ImageJ on the basis of the IF staining of EphB3 in C. E The frequency of Paneth cells distanced from the crypt base was quantified using ImageJ on the basis of the IF staining of lysozyme in C. Values are means ± SD. Statistical analyses were performed by one-way ANOVA with Tukey’s multiple comparisons test (B, D). *p < 0.05, **p < 0.01, ***p < 0.001