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. 2023 Aug 10;14:1174140. doi: 10.3389/fimmu.2023.1174140

Figure 3.

Figure 3

Prophylactic HD5 treatment attenuates radiation-induced colonic mucosal injury. Adult mice were fed a liquid diet with vehicle (Veh-Sham & Veh-IR) or HD5 (HD5-Sham & HD5-IR) for 24 hours before sham-treatment (Sham) or irradiated (IR). TJ and AJ integrity, mucosal permeability, and mucosal inflammatory responses were analyzed 24 hours after irradiation. (A, B) TJ integrity was assessed by immunofluorescence staining of colon cryosections for occludin and ZO-1 (green, occludin; red, ZO-1; blue, nucleus) and confocal microscopy. ZO-1 fluorescence density values are presented in panel (B). (C, D) AJ integrity was assessed by staining colon sections for E-cadherin and β-catenin (green, E-cadherin; red, β-catenin; blue, nucleus). β-catenin fluorescence density values are presented in panel (D). (E, F) Mucosal permeability in vivo was evaluated in the colon (E) and ileum (F) by measuring the vascular-to-luminal flux of FITC-inulin. Values are mean ± sem (n = 6); *p<0.05, **p<0.01, ***p<0.001, and ****p<0.0001 for significant difference between the indicated groups; “ns”, not significant. (G-L) At 24 hours after irradiation, total RNA prepared from the colon were analyzed for IL-1β (G), IL-6 (H), TNFα (I), Mcp1 (J), Cxcl1 (K), and Cxcl2 (L). Values are mean ± sem (n = 6); *p<0.05, **p<0.01, and ****p<0.0001 for significant difference between the indicated groups; “ns”, not significant.