Neutrophils Compromise Retinal Pigment Epithelial Barrier Integrity

Figure 2

Neutrophils induced disruption of the tight junction protein ZO-1 in the RPE monolayer. (a) ZO-1 immunofluorescence staining on RPE-Choroidal explants. (b) Quantative analysis of ZO-1 immunofluorecence area on RPE–Choroidal explants. Bovine RPE-Choroidal explants were incubated with neutrophils for 1 to 3 hours at 37^∘C. ZO-1 expression was determined by confocal immunomicroscopy using anti-ZO-1 Ab. Quantitation of ZO-1 staining positive area was analyzed by using LSM 510 image software. *represents 𝑃<.01. Bar=50𝜇m. (c) Western blot analysis of ZO-1 expression after RPE choroidal explants exposed to neutrophils for 3 hours. In Figure 2(c), the result of densitometry from the western blot (upper panel), normalized to GAPDH, shows significant loss of ZO-1 protein, 𝑃<.05.

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