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Figure 4: C5a does not alter zonula occludens-1 levels in endothelial cells exposed to oxygen-glucose deprivation. Representative Western blots of zonula occludens-1 protein are shown. b.End. 3 cells exposed to 2 h oxygen-glucose deprivation failed to show any changes in zonula occludens-1 protein, whereas 4 h oxygen-glucose deprivation led to marked decreases (a). Blot shows three replicates of each condition plus a maker lane (center). A Western blot of actin protein is shown as a housekeeping control. Endothelial cell cultures were then exposed to 2 h oxygen-glucose deprivation , and C5a was added to some oxygen-glucose deprivation exposed cultures at a concentration of 100 nM (C5a). A representative Western blot shows that the addition of C5a failed to change zonula occludens-1 expression compared to uninjured controls (b). Quantification of Western blots from 3 samples collected from 3 separate experiments (9 observations total) is shown in the graph (c). Relative optical densities from the blots are shown normalized to the OD of the actin proteins. OGD: Oxygen-glucose deprivation, ZO: Zonula occludens, ODs: Optical densities, Con: Controls

Figure 4: C5a does not alter zonula occludens-1 levels in endothelial cells exposed to oxygen-glucose deprivation. Representative Western blots of zonula occludens-1 protein are shown. b.End. 3 cells exposed to 2 h oxygen-glucose deprivation failed to show any changes in zonula occludens-1 protein, whereas 4 h oxygen-glucose deprivation led to marked decreases (a). Blot shows three replicates of each condition plus a maker lane (center). A Western blot of actin protein is shown as a housekeeping control. Endothelial cell cultures were then exposed to 2 h oxygen-glucose deprivation , and C5a was added to some oxygen-glucose deprivation exposed cultures at a concentration of 100 nM (C5a). A representative Western blot shows that the addition of C5a failed to change zonula occludens-1 expression compared to uninjured controls (b). Quantification of Western blots from 3 samples collected from 3 separate experiments (9 observations total) is shown in the graph (c). Relative optical densities from the blots are shown normalized to the OD of the actin proteins. OGD: Oxygen-glucose deprivation, ZO: Zonula occludens, ODs: Optical densities, Con: Controls