Fig. 3

Systemic ATAC administration did not alter NF-κB activation in RPE. a The amount of ATAC in blood was measured after 40 days of drug administration in 7.5-month-old (1.98 μg/500 μL) and in 11.5-month-old (3.5 μg/500 μL) animals. Animals treated with ATAC showed significantly higher ATAC blood concentrations than age-matched controls without ATAC treatment at both ages (Mann-Whitney, p ≤ 0.05). b The degree of inhibition on total complement activation by ATAC was measured by a CH50 hemolysis assay. Dilutions of the sera containing different levels of ATAC were performed to calculate the half maximal inhibitory concentration (IC₅₀) for total complement activity. Higher IC₅₀ values proportionally reflect higher levels of complement activity. Note that the IC₅₀ levels are lower (i.e., curves shifted to the left) for ATAC-treated animals compared to untreated controls at both ages. c ATAC administration did not affect NF-κB activation in RPE at both ages of 7.5 and 11.5 months (Mann-Whitney, p > 0.05). d Representative micrographs of NF-κB p65 immunoreactivity in RPE/choroid from each group in c. RPE cells that demonstrate NF-κB p65 nuclear translocalization have purple nuclei and are marked by blue arrows. Unlabeled RPE nuclei are counterstained with hematoxylin (blue only) and are marked by black arrows. Scale bar; 10 μm. RPE retinal pigment epithelium, Ch choroid. e, f RPE/choroid tissue lysates from 11.5-month-old rats with ATAC administration contained the same amount of phosphorylated p65 subunit as in rats without ATAC in drinking-water (Mann-Whitney, p > 0.05). The level of phosphorylated p50, however, was extremely low in both groups