Figure 1 | Mucosal Immunology

Figure 1

From: Development of a primary mouse intestinal epithelial cell monolayer culture system to evaluate factors that modulate IgA transcytosis

Figure 1

Developing a transwell system using mouse primary intestinal epithelial cells. (a) Schematic of timeline for Transwell experiments. Wild-type cells were treated with ±10 μM DAPT (N-[N-(3,5-difluorophenacetyl-L-alanyl)]-S-phenylglycine t-butyl ester)±1 μg ml−1 lipopolysaccharide (LPS) and were analyzed on day 3 after seeding. (b) Cells were fixed and paraffin embedded on the transwell membranes. Sections were cut and stained with the following: hematoxylin and eosin (H&E), anti-ZO-1, anti-Villin, and anti-polymeric immunoglobulin receptor (pIgR). Bar=50 μm. Gene expression analysis was performed by quantitative real-time polymerase chain reaction for (c) pIgR, (d) Reg3g, and (e) Villin1 (Vil1). All samples were normalized to glyceraldehyde 3-phosphate dehydrogenase mRNA, and data were presented as fold change relative to untreated (0% conditioned media) cells (mean±s.e.m.; n≥3 per condition). One-way analysis of variance: (c) F=96.02, P<0.0001; (d) F=3.441, P<0.0376; (e) F=1.085, P<0.3762. ***P<0.001 by Bonferroni’s multiple comparison test. (f) Transepithelial electrical resistance was measured on day 3. The (resistance × area) is shown for each condition (mean±s.e.m., n=6 per group). Statistical analysis by Student’s t-test showed no significant difference between the two groups (P<0.4362).

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