Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Highly Sensitive, Flow Cytometry-Based Measurement of Intestinal Permeability in Models of Experimental Colitis

doi: 10.1016/j.jcmgh.2022.10.004

Figure Lengend Snippet: The OVA assay shows high sensitivity and specificity to detect intestinal barrier dysfunction in a mouse model of DSS-induced colitis. ( A ) OVA or FITC-D assay readouts from the plasma samples of nontreated (n = 8) or DSS-treated (n = 8) mice co-administered FITC-D (12 mg/mouse) and OVA (1 mg/mouse). Data presented are representative of at least 3 independent experiments. Statistical significance was determined by 1-way analysis of variance using the Tukey post hoc test. ( B ) Visualization of OVA localization in the distal colon of nontreated ( upper panel ) or DSS-treated for 6 days ( lower panel ) WT mice. Mice were orally gavaged with 1 mg/100 μL OVA and tissues were harvested 3 hours later for histology and stained with the same rabbit anti-OVA antibody used in the OVA assay. The localization of OVA (red) is highlighted with white arrowheads , along with stains for epithelium (E-cadherin, green), and nuclei (DAPI, blue). DAPI, 4′,6-diamidino-2-phenylindole. ∗ P ≤.05, ∗∗ P ≤.01, ∗∗∗ P ≤ .001, ∗∗∗∗ P ≤ .0001.

Article Snippet: For visualizing of OVA crossing the IEC barrier, rabbit anti-OVA polyclonal antibody (#GTX21221, 1:1000; GeneTex) or goat anti-OVA polyclonal antibody (#0855303, 1:100; MP Biomedicals) and mouse monoclonal anti-mouse E-cadherin antibody (#610182, 1:400; BD Transduction Laboratories) were used.

Techniques: Staining