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: Rabbit anti-ovalbumin polyclonal antibody (100 μL, #GTX21221; GeneTex) diluted in FACST buffer (10 μg/mL final concentration) was added into each well and mixed using a multichannel pipette.

Techniques: Clinical Proteomics, Staining