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Fig. 1 | Journal of Neuroinflammation

Fig. 1

From: Age-related increases in amyloid beta and membrane attack complex: evidence of inflammasome activation in the rodent eye

Fig. 1

Age-dependent increases of ocular MAC, Aβ and NF-κB. a Western blot analysis revealed increasing MAC levels in the rat RPE/choroid with age from 2.5 to 11.5 months old (Kruskal-Wallis, p ≤ 0.05). b Western blot analysis showed enhanced accumulation of high-molecular weight Aβ (MW > 95 kDa) in the rat RPE/choroid with age from 2.5 to 11.5 months old (Kruskal-Wallis, p ≤ 0.05). c ELISA measurements showed a dramatic increase in the soluble Aβ levels in the vitreous fluids of the 11.5 months old (599.1 pg/mL) compared to the 6 months old group (7.5 pg/mL) (Mann-Whitney, p ≤ 0.05). d, e Analysis of MAC deposition (d) or Aβ (e) in rat RPE/choroid demonstrated a significant increase in immunoreactivity with increasing age, with data normalized to the younger age group of 7.5 months (Mann-Whitney, p ≤ 0.05). f NF-κB activation in RPE increased with age. The percentage of RPE cells with nuclear labeling of translocated NF-κB p65 subunit was higher in the retina of the 11.5 months old group compared to the 7.5 months old group (Mann-Whitney, p ≤ 0.05). g Representative micrographs of MAC immunoreactivity at both ages (7.5 and 11.5 months) showed MAC deposition on the basal side of RPE and in choroid. Immunoreactivity was processed with VIP, resulting in a purple color (blue arrows) and nuclei counterstained with Methyl Green. Background immunoreactivity (0) and semi-quantitative scoring of + and ++ are given for the following examples: 7.5 months old (+), 11.5 months old (++), and negative control (0). Note the dark brown choroidal melanocytes in the 11.5 months picture appeared surrounded by red haze due to bright field illumination. h Representative micrographs of Aβ immunoreactivity at both ages (7.5 and 11.5 months) showed positive immunolabeling on the basal side of RPE and in choroid. Immunoreactivity was processed with AEC, resulting in a red color (blue arrows) and nuclei counterstained as blue. Examples of semi-quantitative scores are given as follows: 7.5 months old (++), 11.5 months old (+++), and negative control (0). i Representative micrographs of the RPE nuclei from the 11.5 months old group showed more robust NF-κB p65 immunoreactivity than the 7.5 months old group. Positive NF-κB p65 immunolabeling is purple in RPE nuclei and indicated by blue arrows. RPE nuclei devoid of NF-κB p65 are blue and indicated by black arrows. Scale bars; 10 μm. RPE retinal pigment epithelium, Ch choroid

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