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. Author manuscript; available in PMC: 2018 Dec 15.
Published in final edited form as: Mucosal Immunol. 2018 Jun 15;11(5):1363–1374. doi: 10.1038/s41385-018-0043-2

Figure 2. Bves−/− colons display increased permeability and altered tight junction composition.

Figure 2

(A) Intercellular spaces quantified by electron microscopy (Bves−/− vs. WT 44.0 ± 3.3 nm vs. 29.6 ± 1.3 nm, ***P<0.0001, n=100–105 images from 3 mice). Images taken at 42,000x. White arrows label tight junctions and black arrowheads delineate intercellular spaces on the basolateral side. (B) Amount of penetrated FITC-dextran was quantified via Ussing chamber ex vivo permeability studies (Bves−/− vs. WT, 0.020 ± 0.003 vs. 0.003 ± 0.001 µg/mL, *P<0.05, n=4 in each group). (C) Colonoids were isolated and grown as monolayers on transwells, and transepithelial resistance (TER) was measured daily by Ohm meter. TER differences were quantified on each day, but only day 4 and day 5 exhibited significant differences: Day 4 (Bves−/− vs. WT, 371.7 ± 4.8 vs. 522.2 ± 5.7 ohms, **P<0.01, n=6) and day 5 (326.5 ± 6.8 vs. 447.7 ± 8.2 ohms, **P<0.01, n=6). (D) Claudin 7 immunoblot from colonic crypts (left) and quantification (right) (Bves−/− vs. WT, 70% reduction, *P<0.05, n=4). (E) Immunofluorescence of the colonic epithelium shows redistribution of pMLC although pMLC and MLC immunoblot were nonsignificant (n=4). Scale bars are shown in the bottom right and are 100 µM. (F) Tight junction mRNA expression of Bves−/− and WT crypt isolates. Claudin2 (Bves−/− vs. WT, 1.8 fold increase, *P<0.05, n=7–8); JAM-A (Bves−/− vs. WT, 1.6 fold increase, *P<0.05, n=7–8) ; Zo-1 (Bves−/− vs. WT, 1.4 fold increase, *P<0.01, n=7–8); Claudin1, Claudin3, and Claudin11 (P=ns, n=7–8).