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Fig. 3
Effects of PGE2 and commonly utilized IBD medications on barrier function, bacterial containment and LRE function. (A-C) PGE2 induces mucin expression and release in a dose-dependent manner. (A) The images of Alcian Blue staining with treatments of 0.1, 1 and 10?μM PGE2. The quantification of the whole gut region is shown in B, and the red boxes indicate the quantification areas in C. Scale bars: 100?μm. Alcian Blue intensities of the full intestine (B) and the intestinal lumen (C) are shown (N=100 from three clutches). **P<0.01. ***P<0.001. Error bar, mean±s.e.m. (D) Quantification of relative Alcian Blue intensity in a human enteroid differentiated epithelial monolayer with and without 1?μM PGE2 treatment. ***P<0.001. N=3 patients with three biopsies each. (E-G) Experiment design of DSS and PGE2 treatment (E) for mortality assays (F) or ingested pHrodo-labeled E. coli protein intensity (G). *P<0.05. **P<0.01. N=1145 from nine clutches. Error bar, mean±s.e.m. (G) Fluorescent intensity of ingested pHrodo-labeled E. coli proteins 1, 2 and 3?h after removal from DSS. N=240 from three clutches. Error bar, mean±s.e.m. (H) Applying PGE2 (left panels), mesalamine (middle panels) and 6-mercaptopurine (right panels) to the DSS injury model. The relative Neutral Red (top panels) and Cyto-ID (bottom panels) intensities with treatment alone (dark blue), with treatment during DSS injury for 24?h (light green) and with 5?h of treatment after DSS removal (light blue) are reported relative to untreated controls. **P<0.01. ***P<0.001. Neutral-Red–PGE2, N=505 from three clutches. Neutral-Red–mesalamine, N=695 from six clutches. Neutral-Red–6-mercaptopurine, N=705 from three clutches. Cyto-ID–PGE2, N=196 from three clutches. Cyto-ID–mesalamine, N=189 from three clutches. Cyto-ID–6-mercaptopurine, N=213 from three clutches. Error bar, mean±s.e.m.