Neutrophils Compromise Retinal Pigment Epithelial Barrier Integrity

Figure 3

Neutrophils induced disruption of tight junction protein occludin in the RPE monolayer. (a) Occludin immunofluorescence staining on RPE-Choroidal explants. (b) Quantative analysis of occludin immunofluorecence area on RPE–Choroidal explants. Bovine RPE-Choroidal explants were incubated with neutrophils for 1–3 hours at 37^∘C. Occludin expression was determined by confocal immunomicroscopy using anti-ZO-1 Ab. Quantitation of occludin staining positive area was analyzed by using LSM 510 image software. ∗represents 𝑃<.01.Bar=50𝜇m. (c) Western blot shows the effects of Neutrophils on occludin expression after 3-hour incubation. In (c), the result of densitometry from the western blot (upper panel), normalized to GAPDH, shows significant loss of occludin protein, 𝑃<.05.

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