Figure 3
From: Inflammatory cytokines directly disrupt the bovine intestinal epithelial barrier
Cytokine treatment increases bovine intestinal organoid barrier permeability, as measured by FITC Dextran permeability. (A–D) Representative images of bovine intestinal organoids exposed to 4 kDa FITC Dextran following 24 h cytokine treatment acquired using a ×10 objective lens and GFP (470/525 nm ex/em) LED light cube. Scale bars denote 200 µm. (E,F) Representative images of an untreated enteroid following 70 kDa FITC Dextran exposure collected using a ×20 objective lens and brightfield or GFP LED light cube. Scale bars denote 300 µm. (G,H) Luminal FITC intensity normalized to external FITC intensity following 24 h cytokine treatment or two-hour treatment with 2 mM EGTA as a positive control. C1, C2, and C3 indicate individual enteroid lines. Nested ANOVA (α = 0.05) determined a significant effect of treatment was present for both 4 kDa and 70 kDa FITC Dextran (p = 0.0097 and p = 0.039, respectively). TNFα and IFNγ increase 4 kDa FITC Dextran permeability (p = 0.0257, p = 0.0132, respectively) as determined by Holm-Šídák post hoc analysis. EGTA, a positive control, increases FITC Dextran permeability (p < 0.0001) determined by one-tailed Mann–Whitney non-parametric test (α = 0.05). Holm-Šídák post hoc analysis for 70 kDa FITC Dextran displayed trending, but not significant, effects of TNFα and IFNγ treatment (p = 0.052 and p = 0.063, respectively).
